Shu-Lan Zhang

Northeastern University (Shenyang, China), Feng-t’ien, Liaoning, China

Are you Shu-Lan Zhang?

Claim your profile

Publications (24)3.2 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: The aim of the study was to investigate the inhibitory effects of RNA interference on XIAP gene expression of human endometrial carcinoma RL95-2 cell and the cell apoptosis. Specific small interference RNA (siRNA) of XIAP was designed and composed. Transfection of siRNA was conducted in endometrial carcinoma cell line RL95-2. The XIAP gene mRNA was assessed by real-time PCR and the change of XIAP protein was assessed with Western Blotting. The cell proliferation and apoptosis was assessed by MTT and flow cytometry methods. After transfection of siRNA specifically targeting XIAP, the relative fold of mRNA transfection in the specific transfection group was (0.04 ± 0.06) and the relative protein expression was (0.590 ± 0.178), which was significantly decreased when compared with the control group (P < 0.05); the cell growth inhibition rate in the transfection group was (47.86 ± 4.46) %, which was significantly increased when compared to the control group (P < 0.05). In vitro experiment showed that synthetic siRNA could effectively inhibit the transfection and expression of XIAP gene of human endometrial carcinoma RL95-2 cell at the mRNA level and protein level, thus significantly promote the apoptosis of endometrial carcinoma cell. The mechanisms involved in the apoptosis still require further investigation.
    Cell biochemistry and biophysics. 08/2014;
  • [Show abstract] [Hide abstract]
    ABSTRACT: This study was to investigate the effect of miR-200c on regulation of ovarian cancer cell metastasis potential and explore the underlying molecular events. qRT-PCR was used to analyze the level of miR-200c expression in 48 ovarian cancer and 30 normal ovarian tissue samples. pre-miR-200c was used to manipulate miR-200c expression in ovarian cancer cells for detection of changed phenotypes of tumor cells. Bioinformatics analysis was then used to predict target genes of miR-200c and GO and pathway analyses drew the miR-200c-related gene network. Luciferase reporter assay confirmed the target of miR-200c as ZEB2. Western blot was used to detect gene expressions in ovarian cancer cells. Level of miR-200c expression was much higher in ovarian cancer than in normal ovarian tissues, and miR-200c expression was inversely associated with advanced clinical stage and lymph node metastasis of ovarian cancer (p < 0.01). The database search predicted 186 miR-200c-targeting genes, and GO analysis showed that functions of these target genes were enriched in the protein binding and other biological processes. Furthermore, miR-200c expression inhibited ovarian cancer cell ES-2 migration and invasion capacity by suppression of ZEB2 expression (p < 0.01). Overexpression of miR-200c regulated E-cadherin and vimentin expression in ovarian cancer cells. This study demonstrated high miR-200c expression in ovarian cancer tissues and ZEB2 as a targeting gene of miR-200c, which mediated the effects of miR-200c on regulation of ovarian cancer cell migration and invasion capacity and epithelial-to-mesenchymal transition. Thus, targeting of miR-200c or ZEB2 may serve as a potential therapeutic strategy for control of ovarian cancer.
    Medical oncology (Northwood, London, England). 08/2014; 31(8):134.
  • [Show abstract] [Hide abstract]
    ABSTRACT: Objective: To explore the relationship between the presence of HPV-16 DNA and the expression Treg surface marker Foxp3(+), peripheral blood levels of Th17/Treg cell-associated cytokines and explore their roles and significance in cervical cancer progression. Between January 2012 and October 2012 at Shengjing Hospital of China Medical University, a total of 142 HPV16 positive patients were divided into cervical cancer (CC, n = 60), cervical intraepithelial neoplasia (CIN, n = 65) and control group (n = 17). Cervical liquid-based cytological (LBC) samples were collected to detect E2 and E6 genes of HPV type 16 using multiple real-time polymerase chain reaction (PCR). E2/E6 ratio was used to evaluate the physical status of HPV-16 DNA in host cell genome. The SP immunohistochemical method was used to detect the expressions of FOXP3 in cervical lesions. The concentrations of Th17/Treg cell-associated cytokines were detected by enzyme-linked immunosorbent assay (ELISA). Under the same status of HPV16 DNA in vivo, the levels of Foxp3(+), transforming growth factor-β (TGF-β) and interleukin-10 (IL-10) were significantly higher than those of the control group (P < 0.01) while the levels of interleukin-17 (IL-17) and interleukin-21 (IL-21)were significantly lower than those of the control group (P < 0.05) . In the same disease, HPV16 DNA integration rate grew with the increases of Foxp3(+), TGF-β and IL-10 while IL-17 and IL-21 were opposite. In the different status of HPV16 type DNA, the expression of Foxp3(+) was closely correlated with Federation International of Gynecology and Obstetrics (FIGO) stage, histological grade and lymphnode metastasis (P < 0.05) except for age (P > 0.05). Treg cytokines, HPV16 integration rate and severity of cervical lesions are positively correlated while Th17 cytokines show opposite effects. Th17/Treg cell-associated cytokines may play an important role in the occurrence and development of cervical cancer.
    Zhonghua yi xue za zhi 10/2013; 93(37):2957-2960.
  • [Show abstract] [Hide abstract]
    ABSTRACT: The aberrant expression of human epidermal growth factor receptor-2 (HER-2) has been detected in ovarian cancer. However, the role of HER-2 in the development of ovarian cancer has not been sufficiently elucidated. The objective of this study was to determine the role of HER-2 in the apoptosis and metastasis of SKOV-3 ovarian cancer cells. SKOV-3 cells were transfected with three double‑stranded small interfering RNA (siRNA) molecules that target HER-2. Various sequences were synthesized by T7 transcription in vitro to select the most effective HER-2‑silencing siRNA. SKOV-3 cells were examined for growth inhibition using the MTT proliferation assay and apoptosis was assessed using flow cytometry and TUNEL assay. The Matrigel basement memebrane matrix was used to assess invasion and chemotactic mobility, as a model of tumor cell metastasis. Western blot analysis was used to detect the expression of matrix metallopeptidase-9 (MMP-9), E-cadherin, N-cadherin and vimentin. siRNA interference in HER-2 resulted in decreased cell proliferation and invasion and increased apoptosis. Western blot analysis demonstrated a marked increase in E-cadherin and MMP-9 and a reduction in N-cadherin and vimentin protein levels in the SKOV-3 cells. The suppression of HER-2 expression resulted in apoptosis and the inhibition of metastasis of SKOV-3 cells. Therefore, the overexpression of the HER-2 gene can enhance the metastatic potential of SKOV-3 cells by increasing the protein levels of MMP-9. Epithelial-mesenchymal transition may be involved in the HER-2 siRNA-induced invasion and migration of SKOV-3 cells. Taken together, these results suggest that HER-2 functions as an oncogene and may thus be an attractive therapeutic target in SKOV-3 ovarian cancer cells.
    Oncology Reports 12/2012; · 2.30 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: To investigate the association of interferon (IFN) γ gene polymorphisms and risk and prognosis of HPV cervical infection. PCR-ASP was used for detecting IFN-γ rs2430561 polymorphism in 179 HPV positive patients and 328 HPV negative normal controls. The frequency of A allele of 63.7% (228/358) was significantly higher than the frequency of T allele of 36.3% (130/358) in HPV positive group (P = 0.045). The frequencies were 41.3% (74/179) in AA genotype and 14.0% (25/179) in TT genotype, women carrying AA genotype increased the risk of HPV infection compare with those with TT genotype (OR = 1.784, 95% CI: 1.031 - 3.088, P = 0.039). During follow-up, the rate of HPV positive again in AA genotype was 83.8% (62/74), while TT genotype was 20.0% (5/25). In the analysis of Kaplan-Meier, the cumulative HPV negative rates of AA, TA and TT genotype exhibited significantly different (P = 0.008). The cumulative HPV negative rate of AA genotype was the lowest (1.1% - 5.9%). IFN-γ rs2430561 polymorphisms confer the susceptibility to HPV infection. Women with AA genotype exhibited higher risk of infection and inclined to be continuous status and recurrence after HPV infection.
    Zhonghua fu chan ke za zhi 10/2012; 47(10):738-41.
  • [Show abstract] [Hide abstract]
    ABSTRACT: To explore the clinical significance of squamous cell carcinoma antigen (SCC-Ag) in the diagnosis, treatment and prognosis of cervical squamous cell carcinoma. The serum SCC-Ag concentrations were measured for 1195 patients with cervical squamous cell carcinoma, 54 patients with non-squamous cell type cervical cancer, 325 patients with cervical intraepithelial neoplasm and 69 healthy women treated at Gynecology Ward of Shengjing Hospital, China Medical University from August 2008 to October 2010. And the correlations with their clinical pathological features of squamous cell carcinoma were analyzed and the changes in the diagnosis, treatment and prognosis monitored. Serum SCC-Ag in patients with cervical squamous cell carcinoma showed a sensitivity of 62.32% and a specificity of 90.10% with the optimal cutoff point of diagnosis at 1.45 µg/L. The differences of pretreatment serum SCC-Ag levels were statistically significant in clinical stage, histological differentiation, depth of invasion and lymph node metastasis (P < 0.01). The posttreatment serum SCC-Ag levels of 106 patients undergoing radical surgery and 264 patients on chemotherapy significantly decreased and were significant different with their pretreatment levels (P < 0.05). There was no relationship of serum SCC-Ag levels and human papillomavirus (HPV) opportunistic infection (all P > 0.05). The best threshold values of pretreatment serum SCC-Ag concentration for predicting early postoperative cervical lymph node metastasis and prognosis of cervical cancer were 2.15 and 12.1 µg/L respectively. As a relatively specific tumor maker for cervical squamous cell carcinoma, squamous cell carcinoma antigen is correlated with clinicopathological features of cervical squamous cell carcinoma. And it has important clinical reference values in the diagnosis, prognosis, follow-up evaluation and treatment monitoring of cervical squamous cell carcinoma.
    Zhonghua yi xue za zhi 05/2012; 92(19):1330-3.
  • Wei Li, Xin Wu, Ning Wang, Duo Yin, Shu-Lan Zhang
    [Show abstract] [Hide abstract]
    ABSTRACT: Gastrointestinal stromal tumor (GIST) represents the most common intramural mesenchymal tumor of the gastrointestinal tract, but the synchronous occurrence of GIST in the stomach and gynecological cancer is rare. We present a unique case of a 56-year-old female patient who was diagnosed with the synchronous development of GIST and an isolated parenchymal splenic metastasis of ovarian cancer. She underwent a wide local excision of gastric lesions with splenectomy. A morphological (histological and immunohistochemical) study established a spindle-cell type of gastrointestinal tumor that expressed CD117, and a parenchymal recurrence of ovarian papillary serous adenocarcinoma. The patient has remained alive and disease-free for 30 months since the last operation. A small GIST concomitant with an isolated parenchymal splenic metastasis of ovarian cancer is rarely encountered. The coexistence of GIST with other malignancies constitutes an intriguing oncologic model. Surgeons are advised to be alert against possible primary GIST accompanying other neoplasms.
    Chinese medical journal 12/2011; 124(24):4372-5. · 0.90 Impact Factor
  • Li-Li Zhang, Shu-Lan Zhang, Shi-Zhuo Wang
    [Show abstract] [Hide abstract]
    ABSTRACT: To explore the effects of paeoniflorin on the proliferation and apoptosis of HeLa cells. HeLa cells treated with paeoniflorin at different concentrations for different hours were assessed by the method of methyl thiazolyl tetrazolium (MTT). Cell apoptosis rate and cycle change were detected by flow cytometry with annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI). The morphological change of HeLa cells was observed by transmission electron microscope (TEM). The expressions of Bcl-2, Bax and Caspase-3 in HeLa cells induced by paeoniflorin were detected by immunocytochemistry. After treatment with different concentrations of paeoniflorin for different hours, the proliferation of HeLa cells was inhibited in a dose and time-dependent manner with an IC50 value of 5054, 2965, 2459 µg/ml at 24, 48 and 72 hours respectively (P < 0.05). After treated by different concentrations of paeoniflorin (control group, 1000 and 2000 µg/ml), apoptosis was induced in HeLa cells at a rate of 0.94%, 10.94% and 13.95% respectively. Such effects were dose-dependent (P < 0.05) and the proportion of HeLa cells in Sphase was under a rising trend. Typical apoptotic changes of HeLa cells under the exposure to paeoniflorin were observed under TEM. There was a lowered expression of Bcl-2 and an elevated expression of Bax and Caspase-3 genes versus the control group after a 48-hour paeoniflorin treatment (P < 0.05). Paeoniflorin can significantly induce the apoptosis of HeLa cells through a down-regulation of anti-apoptotic gene Bcl-2 and an up-regulation of pro-apoptotic genes Bax and Caspase-3.
    Zhonghua yi xue za zhi 12/2010; 90(47):3371-5.
  • [Show abstract] [Hide abstract]
    ABSTRACT: To investigate the expression and significance of elastin and fibulin-5 in anterior vaginal tissue of women with pelvic organ prolapse (POP). Between November 2006 and June 2008, 68 patients with POP underwent surgical treatment in Shengjing Hospital of China Medical University were enrolled in this study, who were classified into 10 patients with grade I, 21 patients with grade II, 25 patients with grade III and 12 patients with grade IV in accordance with pelvic organ prolapse quantitation (POP-Q). Meanwhile, 18 cases with early cervical cancer at stage of I b were treated by total hysterectomy and bilateral salpingo-oophorectomy, their anterior vaginal tissues were selected as controls. Immunohistochemical staining was performed to detect the expression of elastin and fibulin-5. (1) Elastin and fibulin-5 were mainly expressed at extracellular matrix(ECM). (2) The positive rate of fibulin-5 expression in anterior vaginal wall were 5% (2/37) in grade III/IV and 26% (8/31) in grade I/IV POP patients, which reached statistical difference (P = 0.035). However, no statistical different expression was found between postmenopausal (13%, 8/60) and non-menopausal patients (2/8), vaginal delivery < or =2 (19%, 5/27) and >2 patients (12%, 5/41, P > 0.05). (3) The positive rate of elastin expression in anterior vaginal wall in POP group was 31% (21/68), which was significantly lower than 72% (13/18) of control group (P = 0.002). Among POP group, 19% (7/37) of elastin expression in grade III/IV POP was significantly lower than 45% (14/31) in grade I/II of POP patients. However, no statistical difference was found between postmenopausal (30%, 18/60) and non-menopausal patients (3/8), vaginal delivery < or =2 (26%, 7/27) and >2 patients(34%, 14/41, P > 0.05). (4) In POP group, both positive expression of fibulin-5 and elastin of anterior vaginal wall was in 6 cases, both negative expression of fibulin-5 and elastin was in 43 cases. It was illustrated that elastin and fibulin-5 had an positive relationship (P = 0.031). The decreased expression of elastin and fibulin-5 was correlated with degree of POP, which indicated that elastin and fibulin-5 may play a role in the pathogenesis of POP.
    Zhonghua fu chan ke za zhi 07/2009; 44(7):514-7.
  • [Show abstract] [Hide abstract]
    ABSTRACT: To study the effect of RNA interference (RNAi) targeting E6AP on the proliferation and apoptosis of HeLa cells. HeLa cells were cultured and divided into 3 groups: blank control group, cells transfected with nonsense siRNA (small interference RNA), and cells transfected with specific E6AP siRNA. The expressions of E6AP mRNA and protein were detected by RT-PCR and Western blot before and after the transfection respectively. Cell proliferation was determined by methylthiazolyl tetrazolium (MTT). The cell apoptosis index was assessed by flow cytometry. Upon treatment with E6AP siRNA for 24, 48 and 72 h, the expression level of E6AP mRNA decreased 33%, 72% and 70% than siRNA treated group. The protein expression levels in 48 h and 72 h E6AP siRNA groups decreased 38%, 59% comparing with those of the nonsense siRNA treated group (P < 0.05). The proliferative capacity of cells transfectd with E6AP siRNA was significantly lower than blank control group (F = 101.38, P < 0.05) and siRNA treated group (F = 38.64, P < 0.05). The apoptosis index of HeLa cells treated with E6AP siRNA was significantly higher than that of the nonsense siRNA (F = 41.48, P < 0.05) and the blank control group (F = 86.36, P < 0.05). SiRNA targeting can effectively suppress the expression levels of E6AP mRNA, corresponding with a proliferation inhibition and an enhanced apoptosis of HeLa cells.
    Zhonghua bing li xue za zhi Chinese journal of pathology 12/2008; 37(12):822-5.
  • [Show abstract] [Hide abstract]
    ABSTRACT: To investigate the expression level of Galectin-9 (Gal-9) in the serum of cervical cancer patients and analyze the clinic significance of it. Collected sera from 29 patients with cervical cancer, 45 patients with cervical inter-epithelial lesion (CIN) and 26 normal females. Determination of Gal-9 was performed by ELISA, and in the group of patients with cervical cancer, these results were compared with the serum vascular endothelial growth factor (VEGF) and C-reactive protein (CRP). Then selecting four patients from the invasive cervical cancer group performed Gal-9 retrospective immuno-staining analysis. Gal-9 concentration was 27.15 ng/L in the group of patients with cervical cancer that was significantly higher compared with the patients with CIN (21.20 ng/L) and the normal females (17.43 ng/L), and the Gal-9 concentration was significantly correlated with both VEGF and CRP in cervical cancer group. Two of these patients, who had a higher Gal-9 serum level, shown more intense staining and the other two patients, with lower serum level of Gal-9, had moderate immunostaining, suggesting that at least part of serum Gal-9 might be produced by cervical cancer tissue. Gal-9 might play a role in the progression and invasion of cervical cancer and warrants further study.
    Zhonghua yi xue za zhi 11/2008; 88(39):2783-5.
  • [Show abstract] [Hide abstract]
    ABSTRACT: To investigate the effects of RNA interference (RNAi) targeting human epidermal growth factor receptor 2 (HER-2) gene on the change of chemosensitivity to cisplatin of ovarian carcinoma cells. Methods Three kinds of HER-2 gene targeting siRNA, HER-2 siRNA I-III, were synthesized and the best one (HER-2 siRNA III) was screened. Human ovarian carcinoma cells of the line SKOV3 were cultured randomly divided into 3 groups: HER-2 siRNA III group, transfected with HER-2 siRNA III, non-specific siRNA group, transfected with non-specific siRNA III, and control group, without transfection. Cisplatin of the concentrations of 0, 0.05, 0.2. 0.4, 0.8, 1.0, 2.0, 10, and 20 microg/ml was added into the culture fluid for 24 h. MTT method was used to detect the proliferation rate of the SKOV3 cells. Other SKOV3 cells were divided into 3 groups: siRNA group, transfected with HER-2 siRNA III, cisplatin group, exposed to cisplatin, and HER-2 siRNA III and exposed to cisplatin. Annexin V method and flow cytometry were used to detect the apoptosis of the SKOV3 cells. The HER-2 gene expression was assessed by Western blotting. The chemosensitivity of transfected cells to cisplatin was measured by MTT. Western blotting was used to detect the protein expression of apoptosis related proteins: Bcl-2, surviving, XIAP, and Smac. After exposed to cisplatin, the cell survival rate decreased as the dose of cisplatin increased. The proliferation rate of the SKOV3 cells transfected with HER-2 siRNA III and exposed to 1 microg/ml cisplatin was (58 +/- 5)%, significantly lower than those of the nonspecific siRNA III transfection group [(65 +/- 6)%] and the control group [(68 +/- 3)%, both P < 0.01]. No significant difference in the cell survival rate was found between the control and nonspecific groups (P > 0.05). The apoptosis rates at different time point of the HER-2 siRNA III + cisplatin group were all higher than those of the other 2 groups (all P < 0.01). The protein expression levels of the antiapoptotic proteins Bcl-2, survivin, and XIAP were of the HER-2 siRNA III + cisplatin group were significantly lower than those of the cisplatin group, and the protein expression level of the apoptotic protein Smac of the HER-2 siRNA III + cisplatin group significantly higher than that of the cisplatin group (all P < 0.05). HER-2 siRNA III induces cell apoptosis significantly and increases the sensitivity of the human ovarian carcinoma cells to cisplatin. The mechanism of induction of cell apoptosis by HER-2 siRNA III + cisplatin may be related to the downregulation of Bcl-2, survivin and XIAP protein and the up-regulation of Smac protein.
    Zhonghua yi xue za zhi 05/2008; 88(13):909-13.
  • [Show abstract] [Hide abstract]
    ABSTRACT: Vulvar squamous cell carcinoma (VSCC) accounts for about 80%-90% of female vulvar malignant tumors, but the etiology is still unclear. This study was to identify genetic alteration in VSCC by comparative genomic hybridization (CGH). The genomic imbalance, that is, gains or losses of chromosomes, in 21 cases of vulvar squamous cell carcinoma was detected by CGH. The common chromosomal alterations in VSCC included gains of chromosomes 3q, 8q and 12q, and losses of chromosomes 4p and 3p. There are multiple chromosomal aberrations in VSCC. The amplification of the tumor suppressor genes and loss of the oncogenes on these regions may be involved in the development and progression of VSCC.
    Ai zheng = Aizheng = Chinese journal of cancer 06/2007; 26(6):572-5.
  • [Show abstract] [Hide abstract]
    ABSTRACT: To investigate the expression of K-Cl cotransport 1(KCC1) in cervical cancer tissues and to investigate its role in the onset and progression of cervical cancer. Specimens of uterus were obtained from 60 patients aged 45.89 (25 approximately 73), 40 with cervical cancer, 10 with cervical intraepithelial neoplasia (CIN), and 10 with chronic cervicitis. Semi-quantitative RT-PCR was used to detect the mRNA expression. Western blotting was used to detect the protein expression of KCC1. Immunofluorescence assay was used to detect the location of KCC1 protein. The mRNA expression and protein expression of KCC1 were both significantly higher in the cervical cancer tissue than those in the CIN and chronic cervicitis tissues (all P < 0.05). The levels of KCC1 in the lowly differentiated cancer tissues (at grades G(2) and G(3)) were significantly higher than those in the highly differentiated cancer tissues (at grade G(1), P < 0.05). Western blotting revealed that the protein expression level of KCC1 was significantly higher in the cervical cancer tissues than in the CIN and chronic cervicitis tissues (both P < 0.05) and the protein expression levels of KCC1 in the cancer tissues at G" 2 and G(3) grades were significantly higher than that in the cancer tissues at grade G(1) (both P < 0.05). There were no significant differences in mRNA and protein expression between the early and terminal cervical cancer tissues. There were no significant differences in the mRNA and protein expression of KCC1 between the cervical cancer cases with or without lymph node metastasis. Immunofluorescence assay showed that KCC1 was located in the cellular membrane in all patients and the KCC1 expression level there was significantly higher in the cervical cancer tissues than in the tissues of CIN and chronic cervicitis. The expression of KCC1 is up-regulated in cervical cancer and it may play an important role in the onset and progression of cervical cancer.
    Zhonghua yi xue za zhi 05/2007; 87(17):1156-9.
  • [Show abstract] [Hide abstract]
    ABSTRACT: To study the mechanism of resistance to chemotherapeutic drugs of ovarian carcinoma and to locate drug-resistance related genes. Comparative genomic hybridization (CGH) was performed on the specimens of carcinoma tissue obtained during operation from 24 patients with ovarian carcinoma, 12 being drug-resistant and six serous, so as to investigate the genomic imbalance. In the drug-resistant group the most common chromosomal alterations were the gains of 3q, 8q, 20q, 11q, 17q, 12p and 1q, and the losses of 9p, 5q, 10q, and 3p. Of these changes the most significant finding was the gain of 17q with a frequency of 66.7% in the drug-resistant group and with a frequency of 16.7% in the drug- sensitive group, and the gain of 1q with a frequency of 66.7% in drug-resistant group and with a frequency of 16.7% in drug-sensitive group, the loss of 3p with a frequency of 50.0% in the drug-resistant group and with a frequency of 8.3% in the drug-sensitive group. The gains of 17q and 1q and the loss of 3p are the common genomic alterations in the drug-resistant groups, which may code ovarian carcinoma drug-resistance related genes.
    Zhonghua yi xue za zhi 05/2007; 87(15):1009-12.
  • [Show abstract] [Hide abstract]
    ABSTRACT: To investigate the effects of RNA interference (RNAi) targeting human epithelial growth receptor-2 (HER-2) gene on the invasive and chemotactic ability of SKOV3 cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the positive control. Lipofectamine 2000 mediated transient transfection was conducted to transmit the siRNA into SKOV3 cells. Three pairs of specifically targeted (HER-2 siRNAI, HER-2 siRNAII, HER-2 siRNAIII) sequence were selected in the coding region of HER-2 mRNA. Transfection of HER-2 siRNA was conducted with lipofectamine 2000 in ovarian carcinoma cell line SKOV3. The HER-2 gene expression was assessed by real-time PCR and western blot assays. Changes of invasive and chemotactic capacity of SKOV3 cells were measured by polycarbonates coated with or without matrigal. Western blot results showed that the expression of GAPDH protein was decreased in specifically transfected cells and with the increase of siRNA dose, the expression of GAPDH protein was decreased. GAPDH protein gray value in control group, different doses (0.5, 1.0, 1.5, 2.0 microg) GAPDH siRNA interference groups were 0.6855 +/- 0.0259, 0.5698 +/- 0.0275, 0.4542 +/- 0.0296, 0.3341 +/- 0.0178 and 0.1816 +/- 0.0180, respectively. There was a significant difference in each group (F = 198.126, P < 0.01). Both HER-2 siRNAII and siRNAIII could inhibit the expression of HER-2 protein. There was a significant difference in inhibition of HER-2 expression between the siRNAIII (0.1562 +/- 0.0067), siRNAII (0.2162 +/- 0.1589) groups and the other groups (F = 69.461, P < 0.01). In comparison of the relative expression levels in each dose (0.5, 1.0, 1.5, 2.0 microg) group, the difference was significant (F = 174.53, P < 0.01). The relative expression of HER-2 mRNA in HER-2 siRNA group after the 1(st), 3(rd), 6(th) interference were 0.0506 +/- 0.0017, 0.0266 +/- 0.0011 and 0.0154 +/- 0.0020, respectively. There was a decreasing trend in expression levels at different times (P < 0.01). The invasion and chemotactic capacity of SKOV3 cells were decreased after transfection of HER-2 siRNA into the cell line (F = 53.707, P < 0.01 vs F = 11.361, P < 0.01). HER-2 siRNAIII can silence the mRNA expression in a time and dose dependent manner. HER-2 siRNA can inhibit cell invasion and chemotactic capacity of SKOV3 cells. The application of HER-2 siRNA extends the list of available therapeutic modalities in the treatment of human ovarian cancers.
    Zhonghua fu chan ke za zhi 01/2007; 42(1):48-53.
  • Ling Ouyang, Shu-lan Zhang, Xin Wang
    [Show abstract] [Hide abstract]
    ABSTRACT: To investigate chromosomal gains and losses in association with human papillomavirus status in the vulvar squamous cell carcinomas (VSCC). We analyzed 21 VSCC with comparative genomic hybridization (CGH) and polymerase chain reaction. Each case had different degree of variances, including gains and losses of partial or whole chromosome. On average, each case had 4.9 (102/21) abnormal regions; losses were more than gains, being respectively 2.6 (54/21) and 2.3 (48/21). Main variations were gains on chromosomes 3q (43%, 9/21), 8q (38%, 8/21), 12q (33%, 7/21) and 9p (19%, 4/21), and losses of 4p (52%, 11/21), 3p (43%, 9/21) and 9p (10%, 2/21). Gains of 3q (73%) and 12q (64%) were significantly more common in human papillomavirus (HPV)-positive VSCC compared to HPV-negative VSCC (10%, 0; P < 0.01). Gains on chromosome 8q (70%) were more commonly seen in HPV-negative compared to HPV-positive VSCC (9%, P < 0.01). Chromosomal gains or losses are detected by CGH in VSCC. The data indicate that gains of 3q and 12q coincide with HPV infection in VSCC.
    Zhonghua fu chan ke za zhi 11/2006; 41(10):701-5.
  • [Show abstract] [Hide abstract]
    ABSTRACT: Hepatocyte growth factor (HGF) specifically binds to its receptor c-met, activates a complex set of intracellular pathways, and plays important roles in regulating tumor invasion, metastasis, and angiogenesis. HGF and c-met are overexpressed in ovarian cancer. This study was designed to investigate the role of HGF in ovarian cancer invasion and its signal transduction pathway. HGF-induced invasion of ovarian cancer cell line SKOV3 was analyzed with Boyden chamber invasion assay. The expression changes of c-met and matrix metalloproteinase-9 (MMP-9) in SKOV3 cells before and after treatment with HGF and nuclear factor kappaB (NF-kappaB) inhibitor pyrrolidine dithiocarbamate (PDTC) were measured by real-time reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and flow cytometry. The expression of NF-kappaB in SKOV3 cells was evaluated by immunocytochemistry and Western blot. The invasive cells were significantly more in HGF group than in control group and PDTC plus HGF group (343+/-13 vs. 167+/-11 and 241+/-10, P<0.01). HGF promoted the expression of MMP-9 mRNA by 5.66+/-0.10 folds, but had no effect on c-met mRNA expression; PDTC suppressed the HGF-driven expression of MMP-9 mRNA by 2.75+/-0.80 folds. The positive rates of c-met and MMP-9 were significantly higher in HGF-treated cells than in control cells [(96.6+/-2.0)% vs. (73.3+/-2.4)%, P<0.01; (74.6+/-4.4)% vs. (16.0+/-2.9)%, P<0.01]. The protein levels of c-met and MMP-9 were significantly higher in HGF-treated cells than in control cells (2.84+/-0.18 vs. 1.65+/-0.19, P<0.01; 2.94+/-0.13 vs. 0.54+/-0.18, P<0.01). PDTC significantly suppressed the HGF-driven expression of MMP-9 protein: the positive rate and protein level of MMP-9 were (25.8+/-3.7)% and 0.87+/-0.14 (P<0.05). HGF promoted the expression of NF-kappaB protein in a time-dependent manner. The expression peak appeared 1 h after treatment with HGF (from 0.42+/-0.11 to 1.16+/-0.21, P<0.01). PDTC significantly inhibited the HGF-driven expression of NF-kappaB protein (0.38+/-0.12, P<0.01). HGF might stimulate the invasion of SKOV3 cells by up-regulating the expression of MMP-9 via NF-kappaB pathway.
    Ai zheng = Aizheng = Chinese journal of cancer 06/2006; 25(5):570-5.
  • [Show abstract] [Hide abstract]
    ABSTRACT: To investigate the effects of RNA interference (RNAi) targeting HER-2 gene on the biological traits of human ovarian carcinoma. siRNA specific to HER-2 gene was synthesized according to the sequence in the GenBank. Human ovarian carcinoma cells of the line SKOV-3 were cultured and divided into 3 groups: control group; non-specific group, transfected with non-specific siRNA; and specific group, transfected with specific HER-2 siRNA. On the 5th day after transfection cisplatin was added into the culture fluid. The expression of HER-2 mRNA and the expression of protein both before and after transfection were detected by RT-PCR and Western blotting. The cell apoptosis was assessed by flow cytometry. The chemosensitivity of transfected cells to cisplatin was determined by MTT method. Since the 3rd day after transfection the expression of HER-2 mRNA of the HER-2 siRNA group was suppressed till 10 days later. On the 7th day after transfection the expression rate of HER-2 protein of the HER-2 siRNA group was (25.5 +/- 0.8)%, significantly lower than those of the nonspecific siRNA group and control group, (95.7 +/- 0.8)% and (96.6 +/- 1.2)% respectively (both P < 0.001). On the 9th day after transfection no expression of HER-2 protein was found in the HER-2 siRNA group. The apoptosis rate of SKOV-3 cells increased time-dependently after transfection in the HER-2 siRNA group and reached the peak, (53.2 +/- 1.0)% on the 6th day, significantly higher than those of the non-specific siRNA group and control group, (4.1 +/- 0.3)% and (4.1 +/- 0.3)% respectively (both P < 0.001). After exposure to cisplatin for 24 hours, the survival rate of the HER-2 siRNA group was (58.4 +/- 0.8)%, significantly higher than those of the nonspecific siRNA group and control group, (68.0 +/- 0.6)% and (67.0 +/- 0.3)% respectively (both P < 0.001). siRNA targeting HER-2 synthesized in vitro and transfected into human ovarian carcinoma cells effectively suppresses the HER-2 expression, induces cell apoptosis, and increases the sensitivity to cisplatin of the cells. The successful application of HER-2 siRNA extends the list of available therapeutic modalities in the treatment of human ovarian cancer.
    Zhonghua yi xue za zhi 04/2006; 86(9):619-23.
  • Shu-lan Zhang, Yi Yu, Tao Jiang, Bei Lin, Hong Gao
    [Show abstract] [Hide abstract]
    ABSTRACT: To study the expression and significance of metastasis suppressor gene KiSS-1 and its receptor GPR54 in epithelial ovarian cancer. The expression and their clinical significance of KiSS-1 and GPR54 were evaluated by RT-PCR in 37 patients with epithelial ovarian cancer, 15 patients with borderline epithelial ovarian tumors, 15 patients with epithelial benign tumors and 11 patients with normal ovarian tissues. The positivity and relative contents of KiSS-1 mRNA in patients with epithelial ovarian cancer (68%, 0.82 +/- 0.09) and borderline epithelial ovarian tumors (60%, 0.80 +/- 0.10) were all higher than in patients with epithelial benign tumor (20%, 0.65 +/- 0.10) and normal ovarian tissues (18%, 0.66 +/- 0.06; P < 0.05). The positivity and relative contents of KiSS-1 mRNA in patients with epithelial ovarian cancer correlated with their clinical stage and lymph node metastasis (P < 0.05). There was no difference in the positivity and relative contents of GPR54 mRNA between patients with epithelial ovarian cancer (70%, 0.79 +/- 0.07), borderline epithelial ovarian tumor (67%, 0.76 +/- 0.10), benign epithelial ovarian tumor (60%, 0.73 +/- 0.07) and normal ovarian tissues (45%, 0.78 +/- 0.08), respectively (all P > 0.05). The positivity and relative contents of GPR54 mRNA in patients with epithelial ovarian cancer did not correlate with their clinical stage, histology grade, lymph node metastasis and production of ascites (P > 0.05). KiSS-1 and GPR54 may play an important role in inhibiting the invasion and metastasis of early epithelial ovarian cancer.
    Zhonghua fu chan ke za zhi 11/2005; 40(10):689-92.