[Show abstract][Hide abstract] ABSTRACT: Radical surgery of primary vaginal carcinoma typically involves partial or complete resection of the vagina, and young patients in particular can experience sexual dysfunction after surgery. Vaginoplasty is mandatory for this population, multiple vaginal reconstructive techniques have been reported. Here we attempted to determine whether the peritoneum is a feasible alternative to the sigmoid colon in vaginoplasty performed during radical surgery.
World Journal of Surgical Oncology 09/2014; 12(1):302. · 1.20 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The molecular basis for the normal cervical squamous epithelium advance to cervical intraepithelial neoplasia (CIN I, CIN II, CIN III) and ultimately to invasive carcinoma has not yet been defined. We explored the abnormal expression of ITCH (AIP4) and its degrading substrate Large Tumor Suppressor 1 (LATS1) in CINs and cervical cancers, which might disrupt the normal differentiation of the cervical epithelia and contribute to the tumorigenesis of the cervix.
A series of 110 samples, comprising 24 cases of normal cervical tissues, 20 cases of CIN I, 26 cases of CIN II/ III and 40 cases of squamous cancer of the cervix (SCC) were used for analysis. The expression of ITCH and LATS1 was assessed in the tissues by immunohistochemistry, and statistically analyzed by SPSS13.0.
The increased nuclear and cytoplasmic expression levels of ITCH and the low membrane expression of LATS1 were strongly associated with the malignant transformation of the cervical epithelium and the histological progression of SCC. Moreover, the high nuclear and cytoplasmic expression levels of ITCH were significantly correlated with clinical stage (P=0.036, P=0.003, respectively) and tumor size (P=0.046,P=0.039, respectively); the low membrane expression of LATS1 was significantly correlated with clinical stage (P=0.036)and tumor size (P=0.023). Both the nuclear and cytoplasmic expression levels of ITCH were inversely associated with the membrane expression of LATS1 in cervical tissues (P.
Clinical and investigative medicine. Médecine clinique et experimentale. 01/2014; 37(6):E384-94.
[Show abstract][Hide abstract] ABSTRACT: Aberrant histone acetylation plays an essential role in the neoplastic process via the epigenetic silencing of tumour suppressor genes (TSGs); therefore, the inhibition of histone deacetylases (HDAC) has become a promising target in cancer therapeutics. To investigate the correlation of histone acetylation with clinicopathological features and TSG expression, we examined the expression of acetylated H3 (AcH3), RARβ2, E-cadherin, and β-catenin by immunohistochemistry in 65 cervical squamous cell carcinoma patients. The results revealed that the absence of AcH3 was directly associated with poor histological differentiation and nodal metastasis as well as reduced/negative expression of RARβ2, E-cadherin, and β-catenin in clinical tumour samples. We further demonstrated that the clinically available HDAC inhibitors valproic acid (VPA) and suberoylanilide hydroxamic acid (SAHA), in combination with all-trans retinoic acid (ATRA), can overcome the epigenetic barriers to transcription of RARβ2 in human cervical cancer cells. Chromatin immunoprecipitation analysis showed that the combination treatment increased the enrichment of acetylated histone in the RARβ2-RARE promoter region. In view of these findings, we evaluated the antitumor effects induced by combined VPA and ATRA treatment in a xenograft model implanted with poorly differentiated human squamous cell carcinoma. Notably, VPA restored RARβ2 expression via epigenetic modulation. Additive antitumour effects were produced in tumour xenografts by combining VPA with ATRA treatment. Mechanistically, the combination treatment reactivated the expression of TSGs RARβ2, E-cadherin, P21 (CIP1) , and P53 and reduced the level of p-Stat3. Sequentially, upregulation of involucrin and loricrin, which indicate terminal differentiation, strongly contributed to tumour growth inhibition along with partial apoptosis. In conclusion, targeted therapy with HDAC inhibitors and RARβ2 agonists may represent a novel therapeutic approach for patients with cervical squamous cell carcinoma.
PLoS ONE 11/2013; 8(11):e80657. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Natural killer (NK) cells accumulate at the maternal-fetal interface in large numbers, but their exact roles in successful pregnancy remain poorly defined. Here, we provide evidence that T(H)17 cells and local inflammation can occur at the maternal-fetal interface during natural allogenic pregnancies. We found that decidual NK cells promote immune tolerance and successful pregnancy by dampening inflammatory T(H)17 cells via IFN-γ secreted by the CD56(bright)CD27(+) NK subset. This NK-cell-mediated regulatory response is lost in patients who experience recurrent spontaneous abortions, which results in a prominent T(H)17 response and extensive local inflammation. This local inflammatory response further affects the regulatory function of NK cells, leading to the eventual loss of maternal-fetal tolerance. Thus, our data identify NK cells as key regulatory cells at the maternal-fetal interface by suppressing T(H)17-mediated local inflammation.
Proceedings of the National Academy of Sciences 12/2012; · 9.81 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To report the outcomes of modified laparoscopic extraperitoneal uterine suspension to anterior abdominal wall for uterine prolapse using mesh.
Twenty-two patients with uterovaginal prolapse, stage 2 or greater according to pelvic organ prolapse quantification (POP-Q), and with desire for uterine preservation, underwent modified laparoscopic extraperitoneal uterine suspension to the anterior abdominal wall bilaterally using mesh. The outcomes were measured by POP-Q and quality-of-life questionnaires. Intraoperative or postoperative complications were also observed.
Patient age was 61.4 ± 12 years, and parity was 3.3 ± 1.8. After surgery, there was significant improvement in POP-Q measurements of Ba, Bp, and C (P < 0.001). The objective cure rate at 1 year was 100%. A significant improvement in quality-of-life scores was observed (P < 0.001). There were no major intraoperative or postoperative complications. However, all patients reported postoperative dragging pain at the points of puncture ports where the mesh was fixed to the abdominal wall. The visual analog scale decreased from a mean 3-day score of 2.9-0 at 1-month follow-up.
Modified laparoscopic extraperitoneal uterine suspension to the anterior abdominal wall using mesh is a feasible and effective method for treating uterine prolapse and is easy to perform.
International Urogynecology Journal 01/2012; 23(7):887-91. · 2.17 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Epigenetic silencing of the tumor suppressor gene, RARβ2, through histone deacetylation has been established as an important process of cervical carcinogenesis. This pivotal role has led to the suggestion that a combination of retinoids selective for RARβ2 with histone deacetylase (HDAC) inhibitors may have therapeutic potential. Valproic acid (VPA), a HDAC inhibitor, has a critical role in the regulation of gene expression through histone acetylation and causes transformed cells to undergo growth arrest, differentiation, and apoptosis. Therefore, we hypothesized that the combination of VPA and ATRA could restore RARβ2 expression, thus resulting in enhanced anti-neoplastic activity in cervical cancer. Here, we show that VPA combined with ATRA led to hyperacetylation of histone H3 and a significant alteration of gene expression in cervical cancer cells, including RARβ2 gene expression, which was upregulated 50- to 90-fold. The combination therapy effectively inhibited the growth of cervical cancer cells more than the single agent treatment both in vitro and in vivo. The additive effects were associated with a significant upregulation of p21(CIP1) and p53 as well as a pronounced decrease in p-Stat3. Furthermore, the combined treatment led to cell cycle arrest predominantly at the G1 phase, and it preferentially induced cell differentiation rather than apoptosis in cervical cancer cells. The differentiation program was determined by the presence of E-cadherinmediated adhesion and activation of the PI3K/Akt pathway. Taken together, these results provide new insight into the mechanisms of enhanced antitumor activity of the HDAC inhibitor and ATRA regimen, thus offering a new therapeutic strategy for cervical cancer patients.
Current Molecular Medicine 01/2012; 12(3):342-54. · 3.61 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Although early-stage cervical cancer can be treated by surgery, distant metastases can be life threatening. It has been a challenge to identify reliable biomarkers as indicators of metastasis or poor prognosis. We investigated the prognostic impact of vimentin, E-cadherin, and β-catenin expression measured by immunohistochemistry staining in samples from 135 patients with clinical stage I or II cervical squamous cell cancer and in normal cervical tissues from 55 patients who underwent hysterectomy for reasons other than neoplasia. Down-regulation of E-cadherin and β-catenin was positively related to histologic differentiation (P < .001), metastasis (P < .001), and recurrence (P < .001), whereas up-regulation of vimentin was inversely related to histologic differentiation, metastasis, and recurrence (P < .0001, .020, and .000, respectively). In univariate Cox regression analysis, high expression of E-cadherin or β-catenin was a positive prognostic indicator for overall survival (P < .001 and P < .001, respectively), whereas high expression of vimentin was a negative indicator (P < .001). In multivariate Cox regression analysis, high expression of E-cadherin was a positive prognostic indicator for overall survival (P = .002), whereas high expression of vimentin was a negative indicator (P = .034). The expression of E-cadherin and vimentin was associated with survival, and the 2 proteins were independent prognostic factors in univariate and multivariate analyses. The combination of a decrease of E-cadherin and an increase in vimentin might be a valuable survival indictor in cervical squamous cell cancer.
Human pathology 01/2012; 43(8):1213-20. · 2.81 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Natural killer (NK) cells play important roles in adoptive cellular immunotherapy against certain human cancers. This study aims to establish a new human NK cell line and to study its role for adoptive cancer immunotherapy. Peripheral blood samples were collected from 54 patients to establish the NK cell line. A new human NK cell line, termed as NKG, was established from a Chinese male patient with rapidly progressive non-Hodgkin's lymphoma. NKG cells showed LGL morphology and were phenotypically identified as CD56bright NK cell with CD16−, CD27−, CD3−, αβTCR−, γδTCR−, CD4−, CD8−, CD19−, CD161−, CD45+, CXCR4+, CCR7+, CXCR1−, and CX3CR1−. NKG cells showed high expression of adhesive molecules (CD2, CD58, CD11a, CD54, CD11b, CD11c), an array of activating receptors (NKp30, NKp44, NKp46, NKG2D, NKG2C), and cytolysis-related receptors and molecules (TRAIL, FasL, granzyme B, perforin, IFN-γ). The cytotoxicity of NKG cells against tumor cells was higher than that of the established NK cell lines NK-92, NKL, and YT. NKG cell cytotoxicity depended on the presence of NKG2D and NKp30. When irradiated with 8 Gy, NKG cells were still with high cytotoxicity and activity in vitro and with safety in vivo, but without proliferation. Further, the irradiated NKG cells exhibited strong cytotoxicity against human primary ovarian cancer cells in vitro, and against human ovarian cancer in a mouse xenograft model. The adoptive transfer of NKG cells significantly inhibited the ovarian tumor growth, decreased the mortality rate and prolonged the survival, even in cases of advanced diseases. A number of NKG cells were detected in the ovarian tumor tissues during cell therapy. In use of the new human NK cell line, NKG would a promising cellular candidate for adoptive immunotherapy of human cancer.
[Show abstract][Hide abstract] ABSTRACT: Cornification is a kind of apoptosis of squamous epithelial cell in the upper layer of the epithelial tissue. The basal cells away from cell-cycle embark on terminal differentiation, stratum, then die by apoptosis. The critical elements interference from the normal differentiation of squamous cells will lead to cell malignant transformation. We would like to put forward a hypothesis that initiation the cornification of squamous cancerous cells by promotion cell differentiation and stratum can promote squamous carcinoma cells death. Attempts of induction squamous cancerous cells terminal differentiation will add to our understanding of the connection between keratinocytes cornification and the death of squamous-cell carcinoma.
Medical Hypotheses 08/2011; 77(5):763-4. · 1.15 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Our previous studies showed a down-regulation of GRIM-19 in primary human cervical cancers, and restoration of GRIM-19 induced tumor regression. The induction of tumor suppressor protein p53 ubiquitination and degradation by E6 oncoportein of high risk-HPV through forming a stable complex with E6AP is considered as a critical mechanism for cervical tumor development. The aims of this study were to determine the potential role of GRIM-19 in rescuing p53 protein and inducing cervical cancer cell apoptosis.
The protein levels of GRIM-19 and p53 were detected in normal cervical tissues from 45 patients who underwent hysterectomy for reasons other than neoplasias of either the cervix or endometrium, and cervical cancer tissues from 60 patients with non-metastatic squamous epithelial carcinomas. Coimmunoprecipitation and GST pull-down assay were performed to examine the interaction of GRIM-19 with 18E6 and E6AP in vivo and in vitro respectively. The competition of 18E6 with E6AP in binding GRIM-19 by performing competition pull-down assays was designed to examine the disruption of E6/E6AP complex by GRIM-19. The augment of E6AP ubiquitination by GRIM-19 was detected in vivo and in vitro ubiquitination assay. The effects of GRIM-19-dependent p53 accumulation on cell proliferation, cell cycle, apoptosis were explored by MTT, flow cytometry and transmission electron microscopy respectively. The tumor suppression was detected by xenograft mouse model.
The levels of GRIM-19 and p53 were concurrently down regulated in cervical cancers. The restoration of GRIM-19 can induce ubiquitination and degradation of E6AP, and disrupt the E6/E6AP complex through the interaction of N-terminus of GRIM-19 with both E6 and E6AP, which protected p53 from degradation and promoted cell apoptosis. Tumor xenograft studies also revealed the suppression of p53 degradation in presence of GRIM-19. These data suggest that GRIM-19 can block E6/E6AP complex; and synergistically suppress cervical tumor growth with p53.
PLoS ONE 07/2011; 6(7):e22065. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The identification of developmental stages in natural killer (NK) cells, especially in human NK cells, has lagged for decades. We characterize four novel populations defined by CD11b and CD27, which can represent the distinct stages of human NK cells from different tissues. Nearly all NK cells from peripheral blood are CD11b(+) CD27(-) populations whereas NK cells from cord blood have CD11b(+) CD27(-) and CD11b(+) CD27(+) populations. Interestingly, we have found large CD11b(-) CD27(-) populations of NK cells from deciduas. We also demonstrate that each population could be characterized by unique functional and phenotypic attributes. CD11b(-) CD27(-) NK cells display an immature phenotype and potential for differentiation. CD11b(-) CD27(+) and CD11b(+) CD27(+) NK cells show the best ability to secrete cytokines. CD11b(+) CD27(-) NK cells exhibit high cytolytic function. We demonstrate that human NK cells at different developmental stages have special functions and describe a new model of human NK cell differentiation.
[Show abstract][Hide abstract] ABSTRACT: As a member of the p53 family, p63 is considered to be an important differentiation regulation transcriptional factor, but the roles of p63 in many epithelial tumourigenesis and metastasis processes are still not clear. This study was designed to investigate the expression of p63 and its isoform in normal tissues and squamous cell cancer tissues of uterine cervix, and its significance in cancer cell differentiation.
The expression of p63 was assessed in cervical tissue and cell lines by immunohistochemistry, RT-PCR and Western Blotting. The relationships between p63 protein, various clinico-pathological features, and the differentiation marker involucrin were analyzed.
ΔΝp63α is the predominant isoform expressed in cervical epithelial tissues, and it is decreased in moderately or poorly differentiated cervical squamous carcinoma, as well as in the HeLa, SiHa and C33A cervical cancer cell lines. The expression level of ΔΝp63α was positively correlated with that of involucrin in cervical squamous cancer tissue, and the expression of ΔΝp63α is decreased with the degree of tumour invasion.
The decrease of ΔΝp63α in cervical squamous cell cancer appears to be associated with the tumour progression, and ΔΝp63α may be a sensitive marker for cervical squamous cancer differentiation.
Clinical and investigative medicine. Medecine clinique et experimentale 01/2011; 34(3):E184-91. · 1.15 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: It was well studied that ErbB2 (HER2/p185(her2/neu)) overexpression in human malignant cancers correlates with poor prognosis and chemo-resistance. Although Trastuzumab (Herceptin) has been widely used in patients with ErbB2-overexpressing metastatic breast cancer, many patients either do not respond to Trastuzumab therapy or progress within 1 year of initiating Trastuzumab treatment. Previously, we reported a novel tumor-inhibitory antibody chA21, which recognized ErbB2 extracellular domain with an epitope distinct from other tumor-inhibitory anti-ErbB2 antibodies. Here, we report that chA21 combined with Paclitaxel or Trastuzumab significantly enhances the tumor-inhibition effects on ErbB2-overexpressing breast and ovarian cancer in xenograft mice. Moreover, the study reveals that the effects by chA21 to cause an enhanced inhibition on cancer cell proliferation and angiogenesis was highly associated with the intrinsic ability of chA21 to down-regulate ErbB2 receptor, inhibit downstream MAPK and PI3K-AKT signal transduction and activate natural killer cells. Our findings show that chA21 may represent a unique anti-ErbB2 antibody with potentials as therapeutic candidate alone or combination with other anti-ErbB2 reagents in cancer therapy.
Cancer Immunology and Immunotherapy 11/2010; 60(3):339-48. · 3.64 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Between August 2007 and May 2009, 28 patients with uterovaginal prolapse, stage 2 or greater, and who desired uterine preservation, underwent laparoscopic extraperitoneal uterine suspension to the anterior abdominal wall bilaterally using mesh. The primary outcome was recurrence, which was evaluated using point C. Secondary outcomes were effects on quality of life (Pelvic Floor Distress Inventory [PFDI-20] and Pelvic Floor Impact Questionnaire [PFIQ-7]) and sexual symptom (Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire [PISQ-12]) scores, operative time, blood loss, duration of hospitalization, and adverse events. After surgery, there was significant improvement in all pelvic organ prolapse quantification (POP-Q) measurements. The POP-Q score for point C was significantly farther from the hymen at 6-months and 1-year follow-up compared with the preoperative value (-7.8 and -8.0 vs 2.6, respectively; p < .001). The objective cure rates at 6 months and 1 year were 96.4% and 94.1%, respectively. There were no major intraoperative or postoperative complications. However, all patients reported postoperative dragging pain at the points of puncture ports where the mesh was fixed to the abdominal wall. The mean visual analog scale decreased from a mean (SD) 3-day score of 2.61 (1.26) to 0 at 1 month follow-up. Baseline PISQ-12 score changed significantly compared with the value at 6 months after operation (28.4 [2.7] vs 29.3 [2.9]; p < .001). The PFDI-20 and PFIQ-7 scores at 6 and 12 months after surgery improved significantly compared with the baseline scores (p < .001). The subjective success rates at 6 months and 1 year were 96.4% and 94.1%. respectively. Laparoscopic extraperitoneal uterine suspension to the anterior abdominal wall using mesh is a simple, safe, and effective procedure for treating uterovaginal prolapse. However, further studies of the long-term efficiency and reliability of this technique are needed to evaluate its value.
[Show abstract][Hide abstract] ABSTRACT: Like many other solid tumors, cervical cancer contains a heterogeneous population of cancer cells. Several investigators have identified putative stem cells from solid tumors and cancer cell lines via the capacity to self renew and drive tumor formation. The aim of this study was to identify and characterize a cancer stem-like cell population from primary carcinoma of the cervix uteri. Cervical carcinoma from 19 patients staged I-II following International Federation of Gynecology and Obstetrics (FIGO) criteria were disaggregated and subjected to growth conditions selective for stem cells. Eight of nineteen tumor-derived cultures encompassed stem-like cells capable of self-renewal, extensive proliferation as clonal non-adherent spherical clusters. Cell markers of spheroid were identified as CD44+CK17+. Cell survival assays showed the sphere-forming cells were only 48% inhibited by doxorubicin whereas 78% inhibited by paclitaxel. Chemo-resistance may partly attribute to the exclusive expression of ABC transporter. To investigate the tumorigenicity of these stem-like cells, xenoengraftment of 10(5) dissociated spheroid cells allowed full recapitulation of the original tumor, whereas the same amount of tumor cells without non-adherent spheroid selection remained non-tumorigenic. Stemness properties of these spheroid cells were further established by reverse transcription-PCR and Western blotting, demonstrating the expression of embryonic and adult stemness-related genes (Oct-4, Piwil2, C-myc, Stat3 and Sox2). Based on these findings, we assert that cervical cancer contain a subpopulation of tumor initiating cells with stem-like properties, thus facilitating the approach to therapeutic strategies aimed at eradicating the tumorigenic subpopulation within cervical cancer.
[Show abstract][Hide abstract] ABSTRACT: Cervical cancer is the most common malignant disease responsible for the deaths of a large number of women in the developing world. Although certain strains of human papillomavirus (HPV) have been identified as the cause of this disease, events that lead to formation of malignant tumors are not fully clear. STAT3 is a major oncogenic transcription factor involved in the development and progression of a number of human tumors. However, the mechanisms that result in loss of control over STAT3 activity are not understood. Gene associated with Retinoid-Interferon-induced Mortality-19 (GRIM-19) is a tumor-suppressive protein identified using a genetic technique in the interferon/retinoid-induced cell death pathway. Here, we show that reduction in GRIM-19 protein levels occur in a number of primary human cervical cancers. Consequently, these tumors tend to express a high basal level of STAT3 and its downstream target genes. More importantly, using a surrogate model, we show that restoration of GRIM-19 levels reestablishes the control over STAT3-dependent gene expression and tumor growth in vivo. GRIM-19 suppressed the expression of tumor invasion- and angiogenesis-associated factors to limit tumor growth. This study identifies another major novel molecular pathway inactivated during the development of human cervical cancer.
Journal of interferon & cytokine research: the official journal of the International Society for Interferon and Cytokine Research 08/2009; 29(10):695-703. · 1.63 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Mesenchymal stem cells (MSCs) have been reported to secrete a variety of cytokines and growth factors acting as trophic suppliers, but little is known regarding the effects of conditioned medium (CM) of MSCs isolated from femurs and tibias of mouse on the artificial activation of mouse oocytes and on the developmental competence of the parthenotes. In the current study, we investigated the effect of CM on the events of mouse oocyte activation, namely oscillations of cytosolic calcium concentration ([Ca(2)+]i), meiosis resumption, pronucleus formation, and parthenogenetic development. The surface markers of MSCs were identified with a fluorescence-activated cell sorter. The dynamic changes of the spindle and formation of pronuclei were examined by laser-scanning confocal microscopy. Exposure of cumulus-oocyte complexes to CM for 40 min was optimal for inducing oocyte parthenogenetic activation and evoking [Ca(2)+]i oscillations similar to those evoked by sperm (95 vs 100%; P > 0.05). Parthenogenetically activated oocytes immediately treated with 7.5 microg/mL cytochalasin B (CB), which inhibited spindle rotation and second polar body extrusion, were mostly diploid (93 vs 6%, P < 0.01) while CB-untreated oocytes were mostly haploid (5 vs 83%, P < 0.01). Consequently, the blastocyst rate was higher in the CB-treated than in the CB-untreated oocytes. There was no significant difference in developmental rate between oocytes activated with CM and 7% ethanol (62 vs 62%, P > 0.05), but the developmental competence of the fertilized oocytes was superior to that of the parthenotes (88 vs 62%, P < 0.05). The present results demonstrate that CM can effectively activate mouse oocytes, as judged by the generation of [Ca(2)+]i oscillations, completion of meiosis and parthenogenetic development.
Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas / Sociedade Brasileira de Biofisica ... [et al.] 07/2009; 42(6):506-14. · 1.08 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The overexpression and amplification of HER-2 gene is associated with the malignant biological behavior of ovarian carcinoma and these tumor cells expressing elevated levels of HER-2 appear to be resistant to the cytolysis of NK-92. In this study, we analyzed the cytolysis effects of NK-92 on human ovarian carcinoma cells (SK-OV-3) after inhibition the expression of HER-2 mRNA by siRNA. Human ovarian carcinoma cell line SK-OV-3 was transfected with siRNA-hairpin expression retroviral vector (HER-2/siRNA) designed to target HER-2 mRNA. A negative control was established utilizing a vector lacking the antisense component (HER-2/negative). The expression levels of HER-2 gene in SK-OV-3/siRNA, and SK-OV-3/negative cell lines were evaluated by semi-quantitative RT-PCR and immunohistochemistry. The growth and the early apoptosis of these cells were assayed by MTT and flow cytometry, respectively. The cytotoxicity of NK-92 against target cells was investigated by LDH. SK-OV-3/siRNA and SK-OV-3 cells were injected subcutaneously into BALB/c nude mice respectively and NK-92 cells were intraperitoneally injected to examine the anti-tumor activity in vivo. The stable cell line (SK-OV-3/siRNA) with a persistent silence of HER-2 was established. The inhibited expression of HER-2 gene was exhibited by semi-quantitative RT-PCR and immunohistochemistry. The suppressed proliferation and the elicitation of early apoptosis cells were observed in SK-OV-3/siRNA cell line. NK-92 cell line can efficiently lyse the SK-OV-3/siRNA cells in vitro and significantly inhibit the growth of tumors xenografted with SK-OV-3/siRNA cells. Suppression of HER-2 gene expression using siRNA combined treatment of NK-92 presents a new strategy for NK-92 biological treatment on the HER-2 expression epithelial tumors.
[Show abstract][Hide abstract] ABSTRACT: Mesenchymal stem cells (MSCs) secrete a variety of cytokines and growth factors in addition to self-renewal and multiple forms of differentiation. Some of these secreted bioactive factors could improve meiotic maturation in vitro and subsequent embryo developmental potential. The aim of the present study was to determine whether in vitro maturation (IVM) of mouse oocyte with or without cumulus cells could be improved by contact with conditioned medium (CM) of MSCs as well as the efficiency of CM to support follicular growth and oocyte maturation in the ovarian organ of mice cultured on soft agar. The developmental potential of matured oocyte was assessed by blastocyst formation after in vitro fertilization (IVF). Germinal vesicle stage oocytes with or without cumulus cells were subjected to IVM in either CM, Dulbecco's modified Eagle's medium (DMEM), alpha-minimum essential medium (alpha-MEM) or human tubal fluid (HTF). Approximately 120 oocytes were studied for each medium. CM produced a higher maturation rate (91.2%) than DMEM (54.7%), alpha-MEM (63.5%) and HTF (27.1%). Moreover, CM improved embryo development to blastocyst stage significantly more than DMEM and HTF (85 vs 7% and 41.7%, respectively) but there was no significant difference compared with alpha-MEM (85 vs 80.3%). The behavior of cortical granules of IVM oocytes cultured in CM revealed cytoplasmic maturation. Moreover, CM also supported preantral follicles growth well in organotypic culture on soft agar resulting in the maturation of 60% of them to developmentally competent oocytes. The production of estrogen progressively increased approximately 1-fold every other day during organ culture, while a dramatic 10-fold increase in progesterone was observed 17 h after human chorionic gonadotropin stimulus at the end of culture. Thus, CM is an effective medium for preantral follicle growth, oocyte maturation, and sequential embryo development.
Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas / Sociedade Brasileira de Biofisica ... [et al.] 12/2008; 41(11):978-85. · 1.08 Impact Factor