[show abstract][hide abstract] ABSTRACT: An increasing data indicates that altered microRNAs (miRNAs) participate in the radiation-induced DNA damage response. However, a correlation of mRNA and miRNA profiles across the entire genome and in response to irradiation has not been thoroughly assessed. We analyzed miRNA microarray data collected from HeLa cells after ionizing radiation (IR), quantified the expression profiles of mRNAs and performed comparative analysis of the data sets using target prediction algorithms, Gene Ontology (GO) analysis, pathway analysis, and gene network construction. The results showed that the altered miRNAs were involved in regulation of various cellular functions. miRNA-gene network analyses revealed that miR-186, miR-106b, miR-15a/b, CCND1 and CDK6 played vital role in the cellular radiation response. Using qRT-PCR, we confirmed that twenty-two miRNAs showed differential expression in HeLa cells treated with IR and some of these miRNAs affected cell cycle progression. This study demonstrated that miRNAs influence gene expression in the entire genome during the cellular radiation response and suggested vital pathways for further research.
Chinese Science Bulletin 12/2013; 58(36). · 1.32 Impact Factor
[show abstract][hide abstract] ABSTRACT: ACTH independent macronodular adrenal hyperplasia (AIMAH) is a rare disorder characterized by bilateral macronodular hyperplasia of the adrenal glands and increased cortisol production with subclinical or overt Cushing's syndrome. Although the family clustering of AIMAH is infrequent, we have tried our best to find such a familial affected pedigree with complete clinical information and successfully collect adrenalectomy tissue samples from two members of this family. Using whole exome sequencing and several variant prioritization strategies based on disease network analysis, we identified Endothelin receptor type A (EDNRA) Ser420Thr mutation as a causative mutation of AIMAH. EDNRA is a member of G protein coupled receptor family and is involved in cardiovascular or polycystic kidney disease. Our findings indicate that the mutation of EDNRA at S420T site should be regard as a potential AIMAH causative variation in familial and sporadic affected patients.
[show abstract][hide abstract] ABSTRACT: Cisplatin is a classic chemotherapy agent used for treating human non-small cell lung cancer (NSCLC). However, cisplatin resistance is a challenge against successful clinical use. Glutathione S-transferase P1 (GSTP1) has been reported to contribute to cisplatin resistance in many studies. MicroRNAs (miRNAs) are short non-coding RNAs that are 21-25 nucleotides in length. They play a role in post-transcriptional gene regulation by inducing repression and/or mRNA degradation. Recent studies have shown that miRNAs are responsible for cisplatin resistance. This study aims to determine whether deregulated miRNAs can sensitize human lung adenocarcinoma cells to cisplatin by targeting GSTP1. Real-time RT-PCR revealed that GSTP1 mRNA expression was 2.7 ± 0.38 folds (p=0.039) upregulated in A549/CDDP cells, compared with the parental A549 cells, while miR-513a-3p expression was 0.34 ± 0.03 folds (p=0.023) downregulated. Luciferase activity assay proved that GSTP1 was a target gene of miR-513a-3p, which was confirmed by Western blot analysis. Furthermore, CCK-8 assay showed that overexpression of miR-513a-3p could enhance cisplatin-induced apoptosis in human lung adenocarcinoma cell lines, A549/CDDP and SPC-A-1. In conclusion, our data demonstrated that miR-513a-3p can sensitize human lung adenocarcinoma cells to cisplatin by targeting GSTP1.
Lung cancer (Amsterdam, Netherlands) 06/2012; 77(3):488-94. · 3.14 Impact Factor
[show abstract][hide abstract] ABSTRACT: To summarize our experience with extraperitoneal robot-assisted laparoscopic radical prostatectomy (RLRP).
Twenty patients with confirmed prostate cancer by transrectal needle biopsy but no metastasis detected by radiographic examination underwent extraperitoneal RLRP, including 7 with Gleason score of less than 6, 10 with a score of 7, 2 with a score of 8, and 1 with a score of 9.
The procedures were performed successfully in all the patients. In 4 cases, a postoperative PSA value of more than 0.2 ng/ml at 4 weeks suggested residual tumor, for which maximal androgen block therapy was administered before elective radiotherapy. Sixteen patients were followed up for 10 to 37 months (mean 15.5 months). In the 20 cases, the operation was completed in a mean of 180 min (range 150-230 min), with the mean installation time of 48.5 min (range 40-60 min) and average blood loss of 298 ml (range 80-800 ml). The mean postoperative eating time was 1.7 days (1 to 3 days), the mean bladder catheter time was 10.7 days (7 to 14 days), and the mean hospital stay was 10.7 days (range 7-14 days). No postoperative complications occurred in these cases. Postoperative pathology showed a Gleason score no higher than 6 in 6 cases, 7 in 5 cases, and no less than 8 in 9 cases.
The technique of extraperitoneal RLRP can be easily mastered by the surgeons and is especially advantageous for complicated pelvic operations.
Nan fang yi ke da xue xue bao = Journal of Southern Medical University 05/2012; 32(5):749-51.
[show abstract][hide abstract] ABSTRACT: microRNAs (miRNAs) are a versatile class of non-coding RNAs involved in regulation of various biological processes. miRNA-122 (miR-122) is specifically and abundantly expressed in human liver. In this study, we employed 3'-end biotinylated synthetic miR-122 to identify its targets based on affinity purification. Quantitative RT-PCR analysis of the affinity purified RNAs demonstrated a specific enrichment of several known miR-122 targets such as CAT-1 (also called SLC7A1), ADAM17 and BCL-w. Using microarray analysis of affinity purified RNAs, we also discovered many candidate target genes of miR-122. Among these candidates, we confirmed that protein kinase, interferon-inducible double-stranded RNA-dependent activator (PRKRA), a Dicer-interacting protein, is a direct target gene of miR-122. miRNA quantitative-RT-PCR results indicated that miR-122 and small interfering RNA against PRKRA may facilitate the accumulation of newly synthesized miRNAs but did not detectably affect endogenous miRNAs levels. Our findings will lead to further understanding of multiple functions of this hepato-specific miRNA. We conclude that miR-122 could repress PRKRA expression and facilitate accumulation of newly synthesized miRNAs.
Nucleic Acids Research 09/2011; 40(2):884-91. · 8.28 Impact Factor
[show abstract][hide abstract] ABSTRACT: Laparoscopic adrenalectomy has become the gold-standard for the surgical treatment of most adrenal lesions. This study evaluated the operative outcome of laparoendoscopic single-site (LESS) retroperitoneoscopic adrenalectomy (LESS-ARA) in comparison with the current standard operation procedure.
Between June and December 2009, 19 patients underwent LESS-ARA, and their outcomes were compared with a contemporary 1:2 matched-pair cohort of 38 patients who underwent standard ARA by the same surgeon. In LESS-ARA, a multichannel port was inserted through a 2.5- to 3.0-cm transverse skin incision below the tip of the 12th rib. The LESS-ARA procedure was performed using a 5-mm 30º laparoscopic camera and two standard laparoscopic instruments. The following parameters were compared between the two groups: demographics, details of the surgery, perioperative complications, postoperative visual analog pain scale score, analgesic requirement, and short-term measures of convalescence.
The finding showed that LESS-ARA and standard ARA were comparable in terms of the estimated blood loss (30 vs 17.5 ml; p=0.64), postoperative hospital stay (6 vs 6 days; p=0.67), and postoperative complications (2 vs 3 patients; p=1.00) for patients with similar baseline demographics and median tumor size (2.1 vs 3.0; p=0.18) cm. The intraoperative hemodynamic values were similar in the two groups. The LESS-ARA group had a longer median operative time (55 vs 41.5 min; p=0.0004), whereas the in-hospital use of analgesics was significantly less (5 vs 12 morphine equivalents; p=0.03).
The LESS retroperitoneoscopic adrenalectomy approach is feasible and offers a superior cosmetic outcome and better pain control, with perioperative outcomes and short-term measures of convalescence similar to those of the standard approach, albeit with a longer operative time.
[show abstract][hide abstract] ABSTRACT: Ionizing radiation (IR) causes severe cellular damage both directly and indirectly and disrupts RNA integrity. RNA strand
breaks are the most frequent type of damage caused by IR. RNA damage is involved in the development of degenerative diseases,
including Alzheimer’s disease and Parkinson’s disease. However, the mechanism of mRNA damage and any resulting pathophysiological
outcomes are poorly understood. This is partly because there is a lack of sensitive tools to monitor damage randomly occurring
in RNA, especially RNA strand break damage in a given RNA. In this work, a method using the reverse transcription polymerase
chain reaction (RT-PCR) after poly(A) addition to 3′-end of RNA to determine RNA strand break damage in a specific RNA by
poly(A) polymerase has been developed. The levels of damage in specific mRNAs, including ABL1, TP53, GADD45A and ATR from IR-treated HeLa cells were examined. Strand breaks were detected in all mRNAs examined. The study provides a novel and
sensitive method based on 3′-end poly(A)-tailing RT-PCR to monitor RNA strand break damage.
KeywordsRNA damage–RNA strand break–ionizing radiation–poly(A) polymerase–RT-PCR
Chinese Science Bulletin 01/2011; 56(30):3172-3177. · 1.32 Impact Factor
[show abstract][hide abstract] ABSTRACT: Recently, some miRNAs have been reported to be connected closely with the development of human hepatocellular carcinoma. However, the functions of these miRNAs in HCC remain largely undefined.
The expression profiles of miR-193b were compared between HCC tissues and adjacent normal liver tissues using qRT-PCR method. This method was also be used to screen the potential target genes of miR-193b. A luciferase reporter assay was conducted to confirm target association. Finally, the functional effect of miR-193b in hepatoma cells was examined further.
miR-193b was significantly down-regulated in most of the HCC tissues compared to the matching non-tumoural liver tissues. Furthermore, ectopic expression of miR-193b dramatically suppressed the ability of hepatoma cells to form colonies in vitro and to develop tumours in nude mice. CCND1 and ETS1 were revealed to be regulated by miR-193b directly. By regulating the expressions of these oncogenes, miR-193b induced cell cycle arrest and inhibited the invasion and migration of hepatoma cells.
miR-193b may function as a tumour suppressor in the development of HCC by acting on multiple tumourigenic pathways.
European journal of cancer (Oxford, England: 1990) 10/2010; 46(15):2828-36. · 4.12 Impact Factor
[show abstract][hide abstract] ABSTRACT: To analyze the influence of benign prostatic hyperplasia (BPH) drugs on incidence and pathology grading of prostate cancer in China.
Retrospectively investigated the history of drug treatment in 1029 cases of BPH in patients from February 1998 to December 2004. According to the history of drug use, the patients were divided into 4 groups: finasteride group, alpha-receptor inhibitor group, finasteride and alpha-receptor inhibitor combination group and control group (untreated group). We gathered pathology sections of patients in all groups, and gave Gleason Score to each. The difference of incidence and pathology grading of prostate cancer were analyzed by Stata 7.0.
The incidence of prostate cancer in the population of our study was 13.5%; The incidence in finasteride group, alpha-receptor inhibitor group, combination group and control group was 9.8%, 16.0%, 10.3% and 18.6%, respectively. There was significant difference between the two groups with the use of finasteride and the two groups without it (P < 0.05). In our study, the ratio of middle or high level pathology grading (Gleason ≥ 7) in prostate cancer patients was 58.3%, the ratio of middle or high level pathology grading prostate cancer patients in the four groups was 71.4%, 59.6%, 67.7% and 40.0%, respectively. In the comparison of composition ratio of middle or high level prostate cancer, there was significant difference between the two groups with the use of finasteride and the two groups without it (P < 0.05).
Finasteride can lower the risk of prostate cancer, but increase the pathology grade of the prostate cancer which has occurred in the same time. The alpha-receptor inhibitor does not have the same effect.
Zhonghua wai ke za zhi [Chinese journal of surgery] 05/2010; 48(10):761-3.
[show abstract][hide abstract] ABSTRACT: In recent years, some miRNAs have been reported to be connected closely with the development of human hepatocellular carcinoma. In our previous studies, a set of miRNAs were revealed to be dysregulated in HCC tissues. However, the functions of these miRNAs in HCC remain largely undefined.
The expression profiles of miR-183 were compared between HCC tissues and adjacent normal liver tissues using qRT-PCR method. This method was used to screen the potential target genes of miR-183. A luciferase reporter assay was conducted to confirm target association. Finally, the functional effect of miR-183 in hepatoma cells was examined.
Among the 25 HCC samples analyzed, microRNA-183 was significantly up-regulated (twofold to 367-fold) in 17 samples compared with the matching nontumoral liver tissues. Programmed cell death 4 (PDCD4) was identified as the target gene of miR-183. Moreover, PDCD4 is a proapoptotic molecule involved in TGF-beta1-induced apoptosis in human HCC cells, we found that miR-183 transfectants were resistant to apoptosis induced by TGF-beta1.
We conclude that miR-183 can inhibit apoptosis in human HCC cells by repressing the PDCD4 expression, and miR-183 may play an important role in HCC development.
[show abstract][hide abstract] ABSTRACT: To present the technique and experience of robotic-assisted laparoscopic radical cystectomy (RARC) by da Vinci surgical system.
From December 2007 to September 2008, 4 patients underwent RARC and urinary diversion. The age of patients was 44 to 63 years old. The body mass index was 22.8 to 27.7. All their clinical stages were lower than T2N0M0. The technique for RARC involving ureters dissection, posterior dissection, lateral pedicle control, anterior dissection, dorsal vein complex control, neurovascular bundles sparing, lymphadenectomy, ureter-ileal anastomosis, urethra-neobladder anastomosis to either ileal conduit urinary diversion or neobladder reconstruction performed extracorporeally.
All the operations were accomplished successfully. The urinary diversion of 2 case was ileal conduit and others was ileal orthophoria neobladder. The operation time was 300 to 450 min. The time of radical cystectomy was 150 to 180 min. The estimated blood loss was 100 to 500 ml. The postoperative hospital stay was 9 to 35 d. The bed rest time was 4 to 9 d. There was 1 patients who had incomplete intestinal obstruction at 8th postoperative day cured by conservative therapy. The patients were followed up for 3 to 12 months, all patients survived without tumor recurrence. The patients have satisfied urinary continence and normal renal functions without hydronephrosis after the operation.
RARC is small incision and safe, the results are definite. It is one of the direction of minimally invasive urologic surgery.
Zhonghua wai ke za zhi [Chinese journal of surgery] 08/2009; 47(16):1242-4.
[show abstract][hide abstract] ABSTRACT: tRNAs play a central role in protein translation, acting as the carrier of amino acids. By cloning microRNAs, we unexpectedly obtained some tRNA fragments generated by tRNA cleavage in the anticodon loop. These tRNA fragments are present in many cell lines and different mouse tissues. In addition, various stress conditions can induce this tRNA cleavage event in mammalian cells. More importantly, angiogenin (ANG), a member of RNase A superfamily, appears to be the nuclease which cleaves tRNAs into tRNA halves in vitro and in vivo. These results imply that angiogenin plays an important physiological role in cell stress response, except for the known function of inducing angiogenesis.
[show abstract][hide abstract] ABSTRACT: MicroRNAs (miRNAs) have recently been proposed as a versatile class of molecules involved in regulation of various biological processes. Although there is emerging evidence that some microRNAs can function as oncogenes or tumor suppressors, the specific role of miRNA in human hepatocellular carcinoma (HCC) is unclear at this point. In this study, we examined the microRNA expression profiles in a set of 20 human HCC specimens by miRNA microarray and quantitative real-time polymerase chain reaction. The results showed that among the 20 HCC samples analyzed, microRNA-101 was significantly down-regulated twofold or more (twofold to 20-fold) in 16 samples compared with the matching nontumoral liver tissues. Using both a luciferase reporter assay and Western blot analysis, we showed that microRNA-101 repressed the expression of v-fos FBJ murine osteosarcoma viral oncogene homolog (FOS) oncogene, a key component of the activator protein-1 (AP-1) transcription factor. Moreover, using a luciferase expression vector (pAP-1-Luc) driven by seven copies of an AP-1 cis-element, we observed that microRNA-101 expression inhibited phorbol 12-myristate 13-acetate (PMA)-induced AP-1 activity. In in vitro Matrigel invasion and Transwell migration assays, enhanced microRNA-101 expression inhibited the invasion and migration of cultured HCC cells, respectively. These findings suggest that microRNA-101 may play an important role in HCC. CONCLUSION: MicroRNA-101, which is aberrantly expressed in HCC, could repress the expression of the FOS oncogene.
[show abstract][hide abstract] ABSTRACT: miRNAs regulate gene expression by inhibiting translation or by targeting messenger RNA (mRNA) for degradation in a post-transcriptional fashion. In the present study, we show that ectopic expression of miR-34a reduces both mRNA and protein levels of cyclin D1 (CCND1) and cyclin-dependent kinase 6 (CDK6). We also demonstrate that miR-34a targets the 3'-untranslated mRNA region of CCND1 as well as CDK6, which in turn interferes with phosphorylation of retinoblastoma. In addition, we show that overexpression of miR-34a induces a significant G1 cell-cycle arrest in the A549 cell line. Taken together, our data suggest that the effects of miR-34a on G1 cell cycle arrest are through the down-regulation of CCND1 and CDK6, which is associated with other targets of miR-34a either additively or synergistically.
[show abstract][hide abstract] ABSTRACT: The initiation and progression of tumor is regulated by multiple genes. Survivin belongs to the inhibitor of apoptosis protein (IAP) family and is overexpressed in most types of human tumors. Apoptin, originally identified from chicken anemia virus (CAV), can specifically induce apoptosis of human tumor cells rather than normal cells. In this study, survivin expression was silenced by microRNA (miRNA)-mediated RNA interference (RNAi); meanwhile, the engineered miRNA vector was also designed to express apoptin gene. The apoptosis and cell growth were then examined by flow cytometry and MTT assay. The miRNA-mediated knockdown of survivin in combination with apoptin overexpression significantly induced apoptosis and inhibited cell growth. Importantly, the combined strategy was more effective on inducing apoptosis and inhibiting cell growth than either survivin downregulation or apoptin overexpression alone. Taken together, the combined strategy offers potential advantages in control of tumorigenesis, and thus deserves further research as a preferred approach in cancer gene therapy.
Cancer biology & therapy 05/2008; 7(7):1053-60. · 3.29 Impact Factor
[show abstract][hide abstract] ABSTRACT: To present our preliminary experience with laparoscopic ureter reimplantation for distal ureteral stricture without an everted ureteral nipple or submucosal tunneling, and to evaluate its clinical feasibility, safety, and validity.
Six patients with distal ureteral stricture underwent transperitoneal laparoscopic ureteral reimplantation. The ureter was reimplanted into the bladder without everting the ureter or tunneling construction. The seromuscular wall of the ureter was anastomosed circumferentially to the detrusor muscle using continuous absorbable sutures.
All patients had no intraoperative complications or need for open conversion. Intravenous urography 3 months later showed normal drainage without obstruction or reflux. In three patients, hydronephrosis was not revealed by ultrasonography 6 months later.
Laparoscopic noneverted ureteral reimplantation is technically simple, reliable, effective, and feasible.
Journal of Endourology 01/2008; 21(12):1505-8. · 2.07 Impact Factor
[show abstract][hide abstract] ABSTRACT: Telomerase expresses in many cancers and may contribute to drug-resistance. The aim of this study is to observe the effect of telomerase reverse transcriptase (hTERT) DNAzyme on growth of A549/DDP cells and to explore the possibility of telomerase as a new target in treatment of drug resistance for lung cancer.
An hTERT DNAzyme was composed. Telomerase activity was measured by telomeric repeat amplification protocol (TRAP) modified from Kim's method. MTT was used to show the influence of hTERT DNAzyme and cisplatin on A549/DDP cells.
The telomerase activity of A549/DDP cells was down-regulated by hTERT DNAzyme in a dose-dependent manner. The growth inhibition rate of A549/DDP cells was 32.9% by hTERT DNAzyme of 0.25μmol/L, and 60.5% by hTERT DNAzyme combined with 3mg/L cisplatin. The CDI of hTERT DNAzyme and cisplatin was 0.9.
hTERT DNAzyme can inhibit the growth of A549/DDP cells and has a synergistic effect with cisplatin. It is suggested that telomerase may be a new target in treatment of drug-resistant lung cancer cells.
Zhongguo fei ai za zhi = Chinese journal of lung cancer 10/2006; 9(5):405-8.
[show abstract][hide abstract] ABSTRACT: The microRNAs (miRNAs) are an extensive class of small noncoding RNAs (18-25 nucleotides) with important roles in the regulation of gene expression. Although a large number of miRNAs have been identified in a variety of eukaryotic systems, the function of the vast majority of these molecules remains unknown. To study the functions of miRNAs, it is crucial to determine their spatial and temporal expression patterns. Although there are some existing methods that can analyze the expression of miRNAs, it is not an easy task for routine gene-expression studies. In this study, we have established a simple method to detect the expression of mature miRNAs. Total RNA was polyadenylated by poly(A) polymerase, and then cDNA was synthesized by a specific reverse transcriptase (RT) primer and reverse transcriptase using the poly(A)-tailed total RNA as templates. The expression of several mature miRNAs was assayed by this method. The expression profile of two miRNAs, determined by the polymerase chain reaction (PCR) assay, was identical to that determined by Northern blotting. All these data show that the poly(A)-tailed RT-PCR is a convenient method to detect the expression of miRNAs.
[show abstract][hide abstract] ABSTRACT: MicroRNAs (miRNAs) are short 20-25 nucleotides RNA molecules that have been shown to regulate gene expressions in a variety of eukaryotic systems. miRNAs are widespread in eukaryotes and several hundred of miRNAs have been identified, but still a lot of miRNAs have not been detected in various eukaryotic organisms. However, it is not an easy work to clone miRNAs by traditional methods. Here, we describe the identification of 27 miRNAs from a human fetal liver cDNA library by a novel cloning method. Low molecular weight RNA fraction (< or = 200 nt) from fetal liver tissue was extracted, and polyadenylated by poly(A) polymerase. A 5' RNA adaptor was ligated to poly(A)-tailed RNA using T4 RNA ligase. After reverse transcription, the cDNA was amplified by PCR with two adaptor primers. The PCR product with a size about 109 bp was recovered and cloned into T vector. After sequencing, database searching, and expression profiling, 5 novel miRNAs were discovered among other 22 known miRNAs in human fetal liver. These finding indicate that a large diverse population of miRNAs may function to regulate gene expression in hepatocyte.