| Background and Objection:Renal cell carcinoma is one of the common malignant tumors of the urinary system, originated in renal tubular epithelial cells. Renal malignancy tumor accounting for adult solid malignancies2%to3%, and kidney of the original malignant tumors accounted for85%of90%. In recent years, the incidence and mortality of kidney cancer was asecended and showed an upward trend. The causes of kidney cancer is not very clear, radiotherapy and chemotherapy ineffective, radical nephrectomy is considered the most effective treatment methods, however, the5-year survival of advanced renal cell carcinoma less than10%. Tumor recurrence and metastasis is still the main factors threatening the lives of patients. The early diagnosis of renal cell carcinoma is more difficult.The lack of early laboratory diagnostic indicators make early detection of early diagnosis and treatment an difficulty issue. The latest studies have shown that there is a close association between kidney cancer development and progression of miRNAs It may provide a new approach for early tumor diagnosis, prognosis and gene therapy.miRNAs are single-stranded RNA consisting of18to25nucleotides.It is widely present in eukaryotic cells. It generates from the DNA transcribing but not translated into proteins, is a non-coding small RNA molecules. miRNA are widely present in eukaryotes and regulate the functioning of other genes in protein synthesis. miRNA genes were about to stand for1%of the entire genome of miRNA through complementary pairing with target RNA to regulate gene expression level after the transfer.It leads to mRNA degradation and translational repression. MiRNA and its target mRNA form a complex regulatory networks, participation, including cell proliferation, apoptosis, cell differentiation, development, stress response, and variety of biological processes.miR-199a is highly conserved miRNA, It expressed abnormally in liver cancer, cervical cancer, prostate cancer and other tumors, and may participate in the development of tumors through the regulation of cell proliferation, apoptosis, migration and invasion force chang. The expression levels may be used as a clinical diagnosis of a variety of tumors, the signs of pathological type and prognosis.The relationship of renal cell carcinoma and mir-199a rarely reported in study.We aimed to explore the mirl99a expression in renal cell carcinoma tissues and cell changes impact on the clinical data of renal cell carcinoma.It may open up a new areas and provide the experimental evidence for the practice and application of the mir-199a in the diagnosis, treatment and early prevention and treatment of kidney cancer.Methods and materials1.Trizol extract tumor and adjacent tissues total RNA.Spectrophotometer detect RNA concentration, purity, agarose gelelectrophoresis, RNA integrity.The real-time quantitative RT-PCR were detected67cases of renal carcinoma tumors and peritumoral normal tissue expression of mir-199a, to analyze the relationship between the different expression with clinicopathological features.2.Use LipofectamineRNAiMAX simulants (Ignorance and carelessness) and miR-199a-5p transient transfection renal cancer cell lines Caki-1. Application of the MTS to detect cell proliferation, growth curves; detected more cell cycle distribution changes, changes in apoptosis by flow cytometry; application of the changes in the Transwell assay cell migration ability.3.Using SPSS17.0statistical processing of experimental results and measurement data, t-test for paired data, count data between the two groups compared using the χ2test of exact probability method, survival analysis, Kaplan-Meier method Log rank test and COX regulation, inspection standards for a=0.05Results:1. Expression of miR-199a of67cases can be detected in both tissues but the miR-199a expression levels in RCC were significantly lower than those in normal tissues, average13.7folds. The expression of miR-199a in renal cell carcinoma T stage and recurrence was negatively correlated (P<0.05).However, miR-199a expression showed no significant difference in age, histological type,lymph node, metastasis and Fuhrman grade (P>0.05). The survival of miR-199a negative group was significantly lower than the positive group.(Log-rank test p=0.019)2. Transfected with miR-199a in renal cell carcinoma line Caki-1, transfected Caki-1-NC cells, used qPCR to detecte miR-199a and NC groups,the results were significantly different. Transfected miR-199a cell proliferation rate was significantly lower and slope of the decline of the growth curve. Flow cytometry detection of miR-199a transfection apoptosis was significantly increased. The proportion of cells in G1phase decreased S phase cell ratio decreased in G2phase cells no significant changes. 3. Transwell experiments showed that transfection of miR-199a, Caki-1cells compared to the blank group and negative control group, significantly reduced the number of cells through the small room. With the control group, negative control group, transfected miR-199a group difference was significant (P=0.002; P=0.001).Conclusion:1. In conclusion of this study, the detection of miR-199a expression levels in renal cell carcinoma was significantly reduced, and of miR-199a expression level of kidney cancer patients the primary tumor infiltration, tumor recurrence and poor prognosis in patients, suggesting that miR-199a may have reference value and may be a potential kidney cancer prognosis indicators for renal cell carcinoma clinical diagnosis and pathologic grading.2. In this study, what is found by vitro-experiments in renal cancer cells over-expression of miR-199a, can lead to cell proliferation was inhibited, the impact of cell cycle G1phase arrest, increased apoptosis of tumor cells; reduces renal cell carcinoma cell filtration capacity. Provides important experimental evidence of miR-199a as a target for cancer gene therapy and anticancer drugs. Background and Objection:DNA Topoisomerase2a (TOP2a) is a gene involved in the proliferation of malignant tumors. The over expression of TOP2a may be associated with transformation and progression of tumors. Its influence in the development and progression of cancer caused increasing interest in its research. This study was aimed to investigate the expression of TOP2a in renal cell carcinoma and its clinical significance.Methods and materials:Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the expression of TOP2a mRNA in36cases of renal cell carcinoma and normal adjacent tissues. The expression of TOP2a was detected in40renal cell carcinoma a19normal adjacent tissues using the immunohistochemical method.Results:Expression of TOP2a can be detected in both tissues. The positive expression rate of TOP2a in carcinoma was significantly higher than that in normal tissues (P<0.05). The expression of TOP2a in renal cell carcinoma and T stage was positively correlated (P<0.05). However, TOP2a expression in carcinoma and normal tissues showed no significant difference in age, gender, lymph node metastasis, metastasis, recurrence (P>0.05)Conclusion:The expression of TOP2a was higher in renal cancer with an expression that was relative to tumor stage. The over-expression of TOP2a may also contribute to the progression of renal cell carcinoma. |
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