本文選題：腎腫瘤 + 腎細胞癌��； 參考：《第二軍醫(yī)大學學報》2016年07期

【摘要】：目的探討RNA干擾p38基因?qū)θ四I癌細胞株786-O增殖、侵襲、細胞周期和細胞對舒尼替尼敏感性的影響。方法構建針對p38的siRNA531和siRNA659兩條siRNA,分別將其轉(zhuǎn)染至腎癌786-O細胞株,即為siRNA531組和siRNA659組,同時設置轉(zhuǎn)染無義siRNA的陰性對照組和僅加轉(zhuǎn)染試劑的空白對照組。應用RT-PCR技術檢測786-O細胞p38mRNA的表達,蛋白質(zhì)印跡法檢測p38蛋白的表達。CCK-8法檢測細胞的增殖情況和對舒尼替尼的敏感性,流式細胞術檢測細胞的周期改變情況,Transwell實驗檢測細胞的侵襲能力。結果 RT-PCR及蛋白質(zhì)印跡法檢測發(fā)現(xiàn)siRNA轉(zhuǎn)染后786-O細胞p38mRNA及蛋白的表達均降低。與空白對照組和陰性對照組相比,siRNA531組和siRNA659組786-O細胞在轉(zhuǎn)染后3~5d時的增殖率均降低(P0.05,P0.01),細胞對舒尼替尼的敏感性增加,兩組對舒尼替尼的IC50值均低于陰性對照組[(3.2±0.3)、(1.4±0.1)μmol/mL vs(5.4±0.2)μmol/mL,P0.05]。siRNA531組、siRNA659組G1期細胞數(shù)量明顯多于對照組,且兩組786-O細胞出現(xiàn)G0/G1阻滯。轉(zhuǎn)染24h后,兩組的穿膜細胞數(shù)分別為56.43±6.02、34.00±8.12,與陰性對照組(76.27±5.08)相比,兩組細胞的侵襲能力均下降(P0.01)。結論通過轉(zhuǎn)染p38特異性siRNA可以成功沉默腎癌細胞株786-O的p38基因的表達,抑制腎癌細胞株786-O的增殖、侵襲能力,增加其對舒尼替尼的敏感性,為后續(xù)研究腎癌治療及靶向耐藥奠定基礎。
[Abstract]:Objective to investigate the effects of RNA interference p38 gene on proliferation, invasion, cell cycle and cell sensitivity of human renal cell carcinoma cell line 786-O. Methods two siRNAs targeting p38 siRNA531 and siRNA659 were constructed and transfected into 786-O cell line of renal cell carcinoma, that is, siRNA531 group and siRNA659 group respectively. The negative control group transfected with nonsense siRNA and the blank control group with only transfection reagent were set up at the same time. The expression of p38mRNA in 786-O cells was detected by RT-PCR, the expression of p38 protein by Western blotting and the proliferation and sensitivity to sunitinib by CCK-8 assay. Cell cycle changes were detected by flow cytometry and the invasion ability was detected by Transwell assay. Results RT-PCR and Western blot analysis showed that the expression of p38mRNA and protein in 786-O cells decreased after siRNA transfection. Compared with the blank control group and the negative control group, the proliferation rate of 786-O cells in the siRNA531 group and the siRNA659 group decreased at 3 and 5 days after transfection, and the sensitivity of the cells to sulnitinib was increased. The IC50 value of sunitinib in both groups was significantly lower than that in negative control group [3.2 鹵0.3 鹵0.3 渭 mol/mL vs(5.4 鹵0.2 渭 mol 路mL 路mL] .siRNA531 group had significantly higher cell number in G1 phase than that in control group, and 786-O cells were blocked by G0/G1 in both groups. 24 hours after transfection, the number of transmembrane cells in the two groups was 56.43 鹵6.02 鹵34.00 鹵8.12, respectively, which was significantly lower than that in the negative control group (76.27 鹵5.08). Conclusion transfection of p38 specific siRNA can successfully silence the expression of p38 gene in 786-O cell line, inhibit the proliferation and invasion of 786-O cell line, and increase its sensitivity to sunitinib. To lay a foundation for further study of renal cell carcinoma treatment and targeted drug resistance.
【作者單位】： 第二軍醫(yī)大學長征醫(yī)院泌尿外科;武警河南總隊醫(yī)院泌尿外科;解放軍458醫(yī)院泌尿外科;第二軍醫(yī)大學長海醫(yī)院泌尿外科;南京大學醫(yī)學院附屬金陵醫(yī)院南京軍區(qū)南京總醫(yī)院泌尿外科;
【基金】：國家自然科學基金(81272817,81172447) 上海市自然科學基金(11ZR1447800)~~
【分類號】：R737.11

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