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目的探讨Beclin 1对蛋白酶体抑制剂抗卵巢癌OVCAR3细胞作用的影响及其机制探讨。方法 2011年10月至2012年4月在中国医科大学附属盛京医院对OVCAR3细胞经不同浓度MG132处理24h后,MTT法检测细胞存活率。空质粒或Beclin 1质粒转染OVCAR3细胞后,不同蛋白酶体抑制剂处理24 h后,MTT法检测细胞存活率。采用sh RNA技术下调OVCAR3细胞的早期自噬关键基因Atg7或采用晚期自噬抑制剂氯喹及巴弗洛霉素A1处理OVCAR3细胞,MTT法检测MG132对细胞的毒性作用。结果与空白对照组比较,1、2、5和10μmol/L浓度的MG132作用24h均能明显抑制OVCAR3细胞的生长,P值分别为0.023、0.005、0.001、0.000,且呈浓度依赖性;与空质粒转染组相比,Beclin 1转染组OVCAR3细胞经硼替佐米、环氧甲酮四肽、乳胞素、MG132四种蛋白酶体抑制剂处理后,细胞存活率差异均有统计学意义下降,P值分别为0.003、0.008、0.006、0.000。与未转染组和空质粒转染组相比,sh Atg7转染组OVCAR3细胞经MG132处理后,细胞存活率差异无统计学意义,P值分别为0.468,0.667。与空白对照组比较,氯喹及巴弗洛霉素A1处理组OVCAR3细胞经MG132处理后,细胞存活率差异无统计学意义,P值分别为0.776,0.802。结论 Beclin 1以不依赖于自噬的方式增加卵巢癌OVCAR3细胞对蛋白酶体抑制剂的敏感性。
Objective To investigate the effect of Beclin 1 on the effect of proteasome inhibitor on ovarian cancer OVCAR3 cells and its mechanism. Methods From October 2011 to April 2012, OVCAR3 cells were treated with different concentrations of MG132 for 24 h in Shengjing Hospital, China Medical University. Cell viability was detected by MTT assay. After transfection of empty plasmid or Beclin 1 plasmid into OVCAR3 cells, the cell viability was detected by MTT assay after different proteasome inhibitors were treated for 24 h. ShcRNA technology was used to down-regulate Atg7, a key early autophagy gene in OVCAR3 cells, and to OVCAR3 cells using chloroquine and bafilomycin A1, a late inhibitor of autophagy. MTT assay was used to detect the cytotoxicity of MG132 cells. Results Compared with the blank control group, MG132 cells treated with 1, 2, 5 and 10 μmol / L of MG132 for 24 h significantly inhibited the growth of OVCAR3 cells, with P values of 0.023, 0.005, 0.001 and 0.000, respectively. Compared with the plasmid transfection group, the cell viability of OVCAR3 cells transfected with Beclin 1 was significantly decreased after treatment with four inhibitors of bortezomib, epimedium tetrapeptide, lactacystin, MG132 proteasome inhibitor , P values were 0.003,0.008,0.006,0.000. Compared with untransfected group and empty plasmid transfected group, there was no significant difference in survival rate of OVCAR3 cells transfected with sh Atg7 by MG132, P values were 0.468,0.667. Compared with the blank control group, the cell viability of OVCAR3 cells treated with chloroquine and bafilomycin A1 was not significantly different after treatment with MG132, P values were 0.776,0.802. Conclusion Beclin 1 increases the sensitivity of ovarian cancer OVCAR3 cells to proteasome inhibitors in an autophagy-independent manner.