Overexpression of LAPTM4B-35 attenuates epirubucin-induced apoptosis of gallbladder carcinoma GBC-SD cells
Overexpression of LAPTM4B-35 attenuates epirubucin-induced apoptosis of gallbladder carcinoma GBC-SD cells
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DOI：
10.1016/j.surg.2010.12.010
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发表时间：
2011-07-01
期刊：
影响因子：
3.8
通讯作者：
Rui, Jing-An
中科院分区：
文献类型：
--
作者：
Zhou, Li;He, Xiao-Dong;Rui, Jing-An
背景资料。已有研究表明，LAPTM4B可促进胆囊癌(GBC)细胞生长，预测GBC预后不良，但LAPTM4B在GBC细胞凋亡中的作用及其机制尚不清楚。将含有完整LAPTM4B和Mock(PcDNA3)开放阅读框的真核表达载体pcDNA3-AE瞬时导入GBC-SD细胞，表阿霉素诱导细胞凋亡。Hoechst 33258染色、PI染色和Annexin V/PI双染流式细胞仪检测细胞凋亡率。免疫印迹法检测蛋白表达。Pc DNA3-AE表达LAPTM4B-35。尽管这些细胞的凋亡率仍高于亲本细胞，但与Mock质粒转染细胞相比，它们的凋亡率明显降低。在与细胞凋亡相关的分子中，与其他细胞相比，pcDNA3-AE转染组细胞中Bcl2和Bcl1表达上调，Bax、Bid和裂解caspase-9、-3表达下调。我们的数据表明，LAPTM4B-35在体外通过线粒体依赖的途径减弱表阿霉素诱导的GBC-SD细胞的凋亡。因此，LAPTM4B-35蛋白可能与GBC耐药有关。(《外科手术》2011；150：25-31。)
Background. It was shown previously that LAPTM4B promoted growth of gallbladder carcinoma (GBC) cells and predicted poor prognosis in GBC; however; its roles and relative mechanisms in apoptosis of GBC cells remain unknown.Methods. The plasmids, pcDNA3-AE, containing the complete open reading frame of LAPTM4B and Mock (pcDNA3), were transfected transiently into GBC-SD cells, followed by induction of apoptosis by epirubicin. Cell apoptosis was determined by Hoechst 33258 staining, propidium iodide (Pi) staining, and Annexin V/PI double staining flow cytometry. Protein expression was detected by immunoblotting.Results. Overexpression of LAPTM4B-35 was observed in cells transfected with pcDNA3-AE. These cells possessed significantly less apoptosis ratios compared with cells transfected with the Mock plasmid, although the values were still greater than those in parent cells. Of the apoptosis-related molecules, expression of Bcl-2 and Bcl-xL was up-regulated in cells transfected with pcDNA3-AE, whereas expressions of Bax, Bid, and cleaved caspase-9 and -3 were down-regulated compared with their expression in other kinds of cells.Conclusion. Our data show that LAPTM4B-35 attenuated epirubicin-induced apoptosis of GBC-SD cells in vitro through a mitochondria-dependent pathway. Therefore, the protein LAPTM4B-35 might be associated with the chemoresistance of GBC. (Surgery 2011;150:25-31.)