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目的:研究雷帕霉素对人胰腺癌细胞SW1990的mTOR信号通路的影响。方法:采用免疫细胞化学证实mTOR信号通路的存在,通过CCK-8法研究雷帕霉素对胰腺癌细胞增殖的影响,通过Western blot和real time PCR分别从蛋白水平和基因水平研究雷帕霉素对mTOR及其下游分子的表达。结果:免疫细胞化学结果显示p-mTOR、p-p70S6K、p-4E-BP1在细胞质中均呈阳性;CCK-8法显示雷帕霉素能明显抑制细胞增殖(P<0.05);Western blot结果显示随着雷帕霉素浓度的增加,p-mTOR、p-p70S6K表达明显减少,而p-4E-BP1蛋白表达明显增加(P<0.05);Real-time PCR结果显示随雷帕霉素浓度的增加,CyclinD1、VEGF、c-myc基因表达明显减少(P<0.05)。结论:人胰腺癌细胞系SW1990中存在mTOR信号通路并处于激活状态;雷帕霉素抑制胰腺癌细胞增殖与雷帕霉素抑制mTOR信号通路活化有关。
Objective: To study the effect of rapamycin on the mTOR signaling pathway of human pancreatic cancer cell line SW1990. Methods: The existence of mTOR signaling pathway was confirmed by immunocytochemistry. The effects of rapamycin on the proliferation of pancreatic cancer cells were studied by CCK-8. The effects of rapamycin on the proliferation of pancreatic cancer cells were analyzed by Western blot and real time PCR. Expression of mTOR and its downstream molecules. Results: Immunocytochemistry showed that p-mTOR, p-p70S6K and p-4E-BP1 were all positive in cytoplasm. CCK-8 assay showed that rapamycin significantly inhibited cell proliferation (P <0.05) The results showed that the expression of p-mTOR and p-p70S6K was significantly decreased and the expression of p-4E-BP1 was significantly increased (P <0.05) with the increase of rapamycin concentration. Real-time PCR showed that the concentration of rapamycin The expression of CyclinD1, VEGF and c-myc were significantly decreased (P <0.05). CONCLUSION: The mTOR signaling pathway is activated and activated in human pancreatic cancer cell line SW1990. The inhibition of rapamycin on pancreatic cancer cell proliferation is related to the inhibition of mTOR signaling pathway by rapamycin.