Studies On The Expression Of MARCCT-1in Renal Cell Carcinoma And Its Effect On Biological Characteristics Of Renal Cell Carcinoma Cell Line A498

Background and objectiveRenal cell carcinoma(RCC) is one of the most common malignant carcinoma which is secondary to bladder tumor in the urinary system cancer. Its high potential of invasiveness leads to a high recurrence and metastasis potential postoperatively and a very adverse prognosis. So the study on the mechanism of growth, recurrence and metastasis and searching for the bio-markers reflecting invasiveness, metastasis and prognosis of RCC patients is urgent in treatment and prognostic guidance. A lot of studies demonstrated that lncRNAs plays an important role on the pathogenesis, metastasis, relapse of cancer, and certain disease-specificity lncRNA were found. Therefore we presumed that lncRNA may also play an crucial role in the invasiveness and metastasis of renal cell carcinoma, and the role of lncRNAs in clear cell renal cell carcinoma(ccRCC) is the focus of the study.Methodes1、We used lncRNAs chip to study two group of primary tumor in renal cell carcinoma,which were divided into two group by distant metastasis categories.2、We used real time PCR method to study the variance of MARCCT-1between ccRCC the expression of MARCCT-1and clinical stages,the pathologic grading and distant metastasis category were also studied.3、We downregulate MARCCT-1in cell line A498by interfered of MARCCT-1siRNA.Then we observed the cell apoptosis, cell proliferative vitality, cell migration and invasiveness activities by CCK-8assay, flow cytometry, transwell migration assay and the wound healing assay respectively.Results1、We firstly selected out320differently expressed lncRNA from ccRCC tumor tissue, in which114lncRNAs were upregulated in distant metastasis group while the left were downregulated.2、We used real time PCR method to study the variance of MARCCT-1and we found that the expression of MARCCT-1in tumor tissue was up-regulated in11samples, down-regulated in3sample compared to the matched adjacent non-tumor tissue. The relative expression content of MARCCT-1in ccRCC is3.52×10-2±5.20×10-2, which was dramatically higher than that of matched adjacent non-tumor tissue with20.106±0.165. According to the pathological grading, the relative expression level of MARCCT-1was7.66±12.19in well differentiated group. and10.29±11.98in the low-moderately differentiated group, which has no significantly diversity. According to the clinical stages, the relative expression level of MARCCT-1was10.53±14.19in I phase, and7.41±9.45in the Ⅱ-Ⅳ phase, which has no significantly diversity. According to distant metastasis categories, the relative expression content of MARCCT-1was21.70±9.24in distant metastasis group, which was significantly higher than that in the non-distant metastasis group with5.50±9.9.3、We downregulate MARCCT-1in cell line A498by interfered of MARCCT-1siRNA,Then we assessed cell proliferation of cell line A498viability by using the CCK-8assay. We found that downregulating MARCCT-1has significant growth inhibition.4、We downregulate MARCCT-1in cell line A498by interfered of MARCCT-1siRNA, Then we observed the cell apoptosis by flow cytometry. We found that downregulating MARCCT-1has no significant difference.5、We downregulate MARCCT-1in cell line A498by interfered of MARCCT-1siRNA.Then we observed the cell migration activity by transwell migration assay. We found that downregulating MARCCT-1has significant cell migration inhibition.6、We downregulate MARCCT-1in cell line A498by interfered of MARCCT-1siRNA.Then we observed the invasiveness activitye by wound healing assay. We found that downregulating MARCCT-1has significant cell invasiveness inhibition.Conclusion1. We discoverd more than300new candidate lncRNAs might be associated with metastasis and invasiveness in ccRCC.2. The expression of MARCCT-1was up-regulated in ccRCC compared with the matched adjacent non-tumor tissues, suggesting that up-regulation of MARCCT-1expression might be associated with the pathogenesis of ccRCC.3. The relative expression of MARCCT-1(tumor tissue/adjacent non-tumor tissues) was up-regulated in distant metastasis group compared with the non-distant metastasis group, suggesting that up-regulation of MARCCT-1expression might be associated with the metastasis and invasiveness of ccRCC. The relative expression level of MARCCT-1has no significant relevance with pathological grading and the clinical stage in ccRCC.4. MARCCT-1siRNA could be effectively transfected to A498cell line, leading the expression level of MARCCT-1down-regulating.5. The down-regulated expression level of MARCCT-1can lead to the inhibition of the proliferation, invasion and migration activities in A498cells.6. MARCCT-1might play the oncogenic activity role in clear cell renal cell carcinoma, and was a potential target in gene therapy of ccRCC.