| BackgroundRenal cell carcinooma (RCC) originated in the proximal tubule epithelial cells, is the most common malignancy of the adult kidney, accounting for adult malignancies2%-3%, which were ranked7and9in the male and female malignancy. There was about270,000new cases worldwide each year, of which120,000died. In China, the incidence of kidney cancer in the urinary tract in second place, and increasing rate of6%per annum over the past20years. Renal cell carcinoma prone to metastasis and is not sensitive to radiotherapy and chemotherapy. Surgical resection remains the best treatment for RCC, but the recurrence rate is about20%-40%. Due to RCC chemotherapy and radiation resistance, and therefore there is no effective treatment after Surgry. In addition, because lack of early detection and subsequent follow-up biomarker, therefore, timely diagnosis and follow-up of renal cancer progress very difficult. In view of this, the level of molecular genetics to explain the occurrence and development mechanisms of RCC, There is an urgent need to solve the problem and looking for help RCC early diagnosis and prediction of postoperative biochemical indicators for early metastasis and potential therapeutic targets.MiRNA expression profiling analysis has confirmed that miR-143in a variety of malignant tumors, such as esophageal cancer, colorectal cancer, bladder cancer, breast cancer, prostate cancer, low expression, suggesting the possible generally associated with tumor in tumorigenesishas an important role. Increase in the expression of miR-143can inhibit tumor cell growth and induce apoptosis, suggesting that their participation in tumor cell proliferation, differentiation, apoptosis and a series of biological processes. In addition, miR-143can increase the sensitivity of gastric and colon cancer cells to chemotherapeutic drugs, and a chemically modified miR-143can be used as anticancer drugs in the treatment of human tumors, suggesting that miR-143may be a potential tumor biomarkers and treatmenttargets. Recent studies have shown that miR-143in head and neck squamous cancer, lung cancer, breast cancer and other tumors can be adjusted by targeting HK2(hexokinase2) gene affect tumor cell growth and metastasis. And also by adjusting the PI3K/Akt and MAPK signaling pathways affect the biological behavior of bladder cancer proliferation, apoptosis, and so on. RCC miRNAs expression profiling analysis found miR-143expression in renal cancer was down regulated and recurrent renal cell carcinoma patients who are more significant level, so speculated that it may be associated with the development of kidney cancer. In view of this the experiment focuses on the miR-143mimics affect biological behavior of renal cell carcinoma cells proliferation and apoptosis. Using target gene prediction software miRecords TargetScan, Pictar, combined with previous literature literature on the miR-143target genes predicted miR-143target genes in renal cell carcinoma may be Bc12, preliminary explore the miR-1432on renal cell carcinoma andmechanism.ObjectiveResearch the function of miR-143in renal cell carcinoma proliferation and apoptosis and its mechanism.MethodsThe miR-143mimics and Negative control transfection sequence were transfected to human renal cell carcinoma cell line A498with Lipo2000liposome,. The experiment was divided into three groups:miR-143mimics transfection group, negative control transfection group, blank control group which was cultured with opti-MEM I serum-free culture medium only. After transfection, the cell proliferation and apoptosis was detected by CCK-8assay and flow cytometry. Used miRecords, TargetScan, Pictar target gene prediction software combined with previous study, we predicted miR-143target genes may Bc12. Using real-time PCR and western-blot validation the effect of miR-143on target genes Bc12expression.ResultAfter transfection of miR-143mimics, as compared with control group, CCK-8test results showed a significant inhibition of proliferation,. Flow cytometry results showed that miR-143mimics transfection increased rate of apotosis (9.10±2.31)%, significantly higher than the negative control group (4.23±1.52)%and blank control group (3.43±1.61)%, the difference among the groups were statistically significant (P <0.05). The real-time PCR and western-blot results showed that, compared with NC-transfected group and blank control group, miR-143mimics transfected group Bc12mRNA expression was no significant difference (P>0.05), Bc12protein expression was significantly reduced (P<0.05).Conclusion1. MiR-143mimics can inhibit renal cell carcinoma cell line A498proliferation and induced apoptosis;2. Its mechanism of action may be achieved by regulating the expression of anti-apoptotic gene Bc12;3. MiR-143mimics is expected to become the new kidney cancer treatment, its clinical application needs further research. |
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