| Objective and background：Renal cell carcinoma (RCC, renal cell carcinoma) is acommon urinary tract tumor, its incidence is increasing, in2010, its incidence accountsfor about3.8%of the adult malignant tumor, accounting for90%95%of the renaltumor. Along with the diagnosis, detection and treatment level, and enhance the earlypart of the prognosis of patients with kidney cancer survival rates, but there are still partof the late shift and renal cell carcinoma patients prognosis is not ideal. Kidney cancercells are not sensitive to chemotherapy and radiotherapy, this problem has become thefocus of today’s kidney cancer treatment.Epidermal growth factor receptor (EGFR) hasbeen confirmed super expressed in the renal cell carcinoma. It is related to the treatmentof EGFR not sensitive to chemotherapy and radiotherapy. It is expected to improve thesituation of RCC not sensitive to chemotherapy and radiotherapy, which brings hope tothe advanced RCC patients.The mechanism of RNAi is the complementary internal orexternal RNA formation of double-stranded RNA induce the gene silencing. RNAi hashigh specificity, high efficiency, high stability, hereditability etc., it has shown greatpotential on the tumor gene research and treatment.The treatment of tumor RNAi tobring a broad prospect, RNAi technology combined radiotherapy for cancer cells toincrease the damage, and side effects of small, for the treatment of tumor opens a newway.This experiment,use RNAi technology to silence the EGFR gene of ACHN RCCcells,which restrains the ACHN RCC cells proliferation, migration and invasion ability,and increases the sensitivity of radiotherapy. It provides a new way for clinicaltreatment of RCC, and it lays theoretical foundation. Methods: Use immunohistochemical and cell immunofluorescence techniques to provethat EGFR are over expression in organization level and cell level. ACHN cells arecultured in vitro，chemical synthesis in EGFR small interference RNA, through theliposome transfection method transfect the siRNA into ACHN kidney cancercells.Detect knockout efficiency of EGFR protein by Western blottechnical.Determinate RCC cell growth curve through the trypan blue staining.withMTT colorimetric analysis test cell proliferation inhibition rate, cell scratchexperimental detects cell migration ability, transwell detects cell invasion ability. Thenrespectively use0,2,4,6,8gy doses of x-rays to radiate5groups of RCC cells (eachgroup contains blank control, negative control, specific RNA interferencegroup),optical microscope to observe cells’ apoptosis, trypan blue staining detects cellapoptosis rate.Results: Tissue and cells levels are seen in high level of EGFR expression in RCC.specific siRNA can obviously inhibit the expression of EGFR.Growth curve hint thatsiRNA group cells has a poor growth, obviously within24h, and h,72h,96hproliferation inhibition rate were15.6%,38.4%,64.1%,68%, are higher than the blankgroup and negative group (P <0.05).Compared with blank group and the negativecontrol group,cell migration and invasion ability of siRNA group are loweredobviously.Optical microscope to observe RNAi adjuvant radiotherapy group cell deathsituation was obviously, trypan blue staining detection from specific RNA interferencegroup can significantly enhance the sensitivity of ACHN RCC cells to radiation therapy(P <0.05).Conclusion: In vitro studies show that proliferation, migration and invasion ability ofRCC cells are significantly reduced after siRNA inhibiting EGFR expression, Silencethe EGFR gene of ACHN RCC cells can obviously improve the cells’ sensitivity of the radiation therapy, it provides a new train of thought for RCC treatment.Use the RNAitechnology to silence EGFR gene is expected to become the new method of kidneycancer gene therapy. |
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