沉默HCCR-2基因对胰腺癌细胞Bcl-2和Bax表达及细胞增殖与凋亡的影响  

Effect on the expression of Bcl-2,Bax and proliferation,apoptosis in pancreatic cancer cells by silencing of the HCCR-2 gene

作　　者：姜琳[1] 孙灿林[1] 黄俊星[2] 林梅[2] 吴正东[2] 崔永安[2] 肖蔚[3] 焦霞[3] 叶军[4] 于鸿[2,3] JIANG Lin;SUN Can-lin;HUANG Jun-xing;LIN Mei;WU Zheng-dong;CUI Yong-an;XIAOWei;JIAO Xia;YE Jun;YU Hong(Department of Anesthesiology,Taizhou People's Hospital,Taizhou 225300,China)

机构地区：[1]泰州市人民医院麻醉科,江苏省225300 [2]泰州市人民医院肿瘤科,江苏省225300 [3]泰州市人民医院病理科,江苏省225300 [4]泰州市人民医院医学研究中心,江苏省225300

出　　处：《中华临床医师杂志（电子版）》2012年第22期7208-7213,共6页Chinese Journal of Clinicians(Electronic Edition)

基　　金：江苏省"333工程"培养资金资助项目(200924);江苏省自然科学基金(BK2010357)

摘　　要：目的利用反义寡核苷酸(ASODN)干扰技术探讨人宫颈癌癌基因-2(HCCR-2)对人胰腺癌细胞增殖、凋亡的影响及其可能的分子机制。方法经脂质体介导将HCCR-2ASODN转染人胰腺癌细胞株PANC-1,荧光显微镜观察细胞内荧光信号,流式细胞仪检测细胞转染效率及凋亡率;噻唑盐(MTT)比色法检测转染细胞的增殖活性;逆转录-聚合酶链反应(RT-PCR)及Western blot检测转染细胞HCCR-2、Bcl-2、Bax mRNA与蛋白表达。结果 HCCR-2ASODN转染PANC-1细胞的转染效率为85.04%。反义组、无义组和单加培养液组细胞凋亡率分别为(24.04±2.38)%、(2.44±0.13)%、(2.54±0.21)%,反义组较其他组细胞凋亡显著增加(P<0.01);相应的各组细胞增殖活性分别为2.38±1.03、3.78±1.69、3.72±1.54,反义组细胞增殖较其他组显著被抑制(P<0.01)。反义组、无义组和单加培养液组细胞HCCR-2mRNA表达量分别为0.29±0.16、0.55±0.31、0.57±0.33;Bcl-2mRNA表达量分别为0.31±0.14、0.63±0.28、0.62±0.35;BaxmRNA表达量分别为0.68±0.36、0.24±0.09、0.23±0.12;HCCR-2蛋白表达量分别为0.47±0.28、0.93±0.48、0.95±0.42;Bcl-2蛋白表达量分别为0.35±0.19、0.82±0.51、0.81±0.45;Bax蛋白表达量分别为0.91±0.42、0.42±0.21、0.39±0.20,较其他组,反义组细胞HCCR-2、Bcl-2mRNA及蛋白表达量显著降低(P<0.01),而Bax mRNA及蛋白表达量显著增加(P<0.01)。结论下调PANC-1细胞HCCR-2表达后,细胞增殖受到抑制,细胞凋亡增加,其作用机制与Bcl-2表达降低而Bax表达升高有关。Objective To evaluate the effect and mechanism of human cervical cancer oncogene-2 oligodeoxynucleotide antisense(HCCR-2 ASODN)on proliferation and apoptosis of pancreatic cancer cells.Methods HCCR-2 ASODN,nonsense oligodeoxynucleotide(NSODN)were transfected into PANC-1 cells.The transfection rate was accessed by fluorescence microscopy and flow cytometry.The cell proliferation and apoptosis were observed by MTT assay and flow cytomertry.The mRNA and protein expressions of HCCR-2,Bcl-2 and Bax were detected by RT-PCR and Western blot.Results HCCR-2 ASODN was successfully transfected into PANC-1 cells.The transfection efficiency of PANC-1 cells was 85.04%.The apoptosis rate in HCCR-2 ASODN,NSODN and control groups were(24.04±2.38)%,(2.44±0.13)%,(2.54±0.21)%.The apoptosis of PANC-1 cells was significantly increased in HCCR-2 ASODN transfected group,compared with the other two groups(P<0.01).The A490 value of cells in HCCR-2 ASODN,NSODN and control groups were 2.38±1.03,3.78±1.69,3.72±1.54.The proliferation of PANC-1 cells was significantly decreased in HCCR-2 ASODN transfected group,compared with the other two groups(P<0.01).The mRNA levels of HCCR-2,Bcl-2 and Bax in HCCR-2 ASODN,NSODN and control groups were:HCCR-2 0.29±0.16,0.55±0.31,0.57±0.33;Bcl-2 0.31±0.14,0.63±0.28,0.62±0.35;Bax 0.68±0.36,0.24±0.09,0.23±0.12.Accordingly,protein levels were:HCCR-2 0.47±0.28,0.93±0.48,0.95±0.42;Bcl-2 0.35±0.19,0.82±0.51,0.81±0.45;Bax 0.91±0.42,0.42±0.21,0.39±0.20.The·8 0 2 7·中华临床医师杂志(电子版)2012年11月第6卷第22期Chin J Clinicians(Electronic Edition),November 15,2012,Vol.6,No.22 expressions of mRNA and protein of HCCR-2,Bcl-2 were significantly reduced,however the expressions of mRNA and protein of Bax were significantly increased in HCCR-2 ASODN transfected group,compared with the other two groups(P<0.01).Conclusions HCCR-2 ASODN transfection causes significant increase in the apoptosis and decreases in the proliferation of PANC-1 cells.These effects are related to the downregul

关 键 词：寡核苷酸类 反义 癌基因 宫颈肿瘤 基因 BCL-2 BCL-2相关X蛋白质 细胞凋亡 

分 类 号：R735.9[医药卫生—肿瘤]