肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2013年
9期
577-580
,共4页
樊丽君%李美宁%牛万通%孙建雯%程牛亮
樊麗君%李美寧%牛萬通%孫建雯%程牛亮
번려군%리미저%우만통%손건문%정우량
结肠肿瘤%激活蛋白2α%奥沙利铂
結腸腫瘤%激活蛋白2α%奧沙利鉑
결장종류%격활단백2α%오사리박
Colonic neoplasms%Activating protein 2α%Oxaliplatin
目的 研究转录因子激活蛋白2α (Ap-2α)在结肠癌HCT-116细胞对奥沙利铂抗药性中的作用及相关机制.方法 利用脂质体介导将Ap-2 α RNA干扰质粒GV102-Ap-2 α-RNAi(实验组)及对照质粒GV102-NC(阴性对照组)转染至HCT-116细胞中；用实时荧光定量聚合酶链反应(PCR)法与Western blot法检测实验组、阴性对照组及未处理的HCT-116空白对照组AP-2α表达情况；用CCK-8法研究HCT-116细胞增殖情况；用PE标记流式细胞术检测细胞凋亡情况.结果 转染载有AP-2 α-RNAi序列的质粒,干扰序列1干扰后Ap-2 α mRNA与蛋白水平均下降最显著.奥沙利铂处理HCT-116细胞后Ap-2α蛋白水平随作用时间延长而升高,但mRNA变化不明显；奥沙利铂作用后实验组Ap-2α蛋白水平低于阴性对照组；CCK-8法结果提示奥沙利铂处理的实验组细胞增殖水平[(88±3)％]高于奥沙利铂处理的阴性对照组[(57±3)％]与空白对照组[(73±4)％],同时流式细胞术结果表明奥沙利铂处理实验组细胞凋亡率[(15.07±1.20)％]低于奥沙利铂处理阴性对照组[(24.93±0.90)％]与空白对照组(23.71％±1.32％).结论 AP-2 α表达降低可能与结肠癌细胞对奥沙利铂的抗药性有关.
目的 研究轉錄因子激活蛋白2α (Ap-2α)在結腸癌HCT-116細胞對奧沙利鉑抗藥性中的作用及相關機製.方法 利用脂質體介導將Ap-2 α RNA榦擾質粒GV102-Ap-2 α-RNAi(實驗組)及對照質粒GV102-NC(陰性對照組)轉染至HCT-116細胞中；用實時熒光定量聚閤酶鏈反應(PCR)法與Western blot法檢測實驗組、陰性對照組及未處理的HCT-116空白對照組AP-2α錶達情況；用CCK-8法研究HCT-116細胞增殖情況；用PE標記流式細胞術檢測細胞凋亡情況.結果 轉染載有AP-2 α-RNAi序列的質粒,榦擾序列1榦擾後Ap-2 α mRNA與蛋白水平均下降最顯著.奧沙利鉑處理HCT-116細胞後Ap-2α蛋白水平隨作用時間延長而升高,但mRNA變化不明顯；奧沙利鉑作用後實驗組Ap-2α蛋白水平低于陰性對照組；CCK-8法結果提示奧沙利鉑處理的實驗組細胞增殖水平[(88±3)％]高于奧沙利鉑處理的陰性對照組[(57±3)％]與空白對照組[(73±4)％],同時流式細胞術結果錶明奧沙利鉑處理實驗組細胞凋亡率[(15.07±1.20)％]低于奧沙利鉑處理陰性對照組[(24.93±0.90)％]與空白對照組(23.71％±1.32％).結論 AP-2 α錶達降低可能與結腸癌細胞對奧沙利鉑的抗藥性有關.
목적 연구전록인자격활단백2α (Ap-2α)재결장암HCT-116세포대오사리박항약성중적작용급상관궤제.방법 이용지질체개도장Ap-2 α RNA간우질립GV102-Ap-2 α-RNAi(실험조)급대조질립GV102-NC(음성대조조)전염지HCT-116세포중；용실시형광정량취합매련반응(PCR)법여Western blot법검측실험조、음성대조조급미처리적HCT-116공백대조조AP-2α표체정황；용CCK-8법연구HCT-116세포증식정황；용PE표기류식세포술검측세포조망정황.결과 전염재유AP-2 α-RNAi서렬적질립,간우서렬1간우후Ap-2 α mRNA여단백수평균하강최현저.오사리박처리HCT-116세포후Ap-2α단백수평수작용시간연장이승고,단mRNA변화불명현；오사리박작용후실험조Ap-2α단백수평저우음성대조조；CCK-8법결과제시오사리박처리적실험조세포증식수평[(88±3)％]고우오사리박처리적음성대조조[(57±3)％]여공백대조조[(73±4)％],동시류식세포술결과표명오사리박처리실험조세포조망솔[(15.07±1.20)％]저우오사리박처리음성대조조[(24.93±0.90)％]여공백대조조(23.71％±1.32％).결론 AP-2 α표체강저가능여결장암세포대오사리박적항약성유관.
Objective To investigate the effect of AP-2α on the chemoresistance to oxaliplatin of colorectal carcinoma cell and its related mechanism.Methods Plasmid of GV102-AP-2α-RNAi (experimental group) and control plasmid GV102-NC (negative control group) were transfected into HCT-116 using Lipofectamine 2000 respectively.The AP-2α expression levels of mRNA and protein of experimental group,control group and HCT-116 blank group were detected by qRT-PCR and Western blot.Cell proliferation assay was performed using the CCK-8 and the apoptosis assays were preformed with Annexin V-PE Apoptosis Kit.Results The AP-2α expression levels of mRNA and protein both decreased after transfection of AP-2α-RNAi plasmid,moreover,the effect produced by subsequence 1 was the most significant.After treatment by oxaliplatin,AP-2α protein levels increased with time while mRNA did not change significantly.Western blot results suggested that the level of AP-2α protein in experimental group which was maintained in oxaliplatin was lower than the negative control group.CCK-8 results suggested that cell proliferation ability was significantly higher for the experimental group maintained in oxaliplatin [(88±3) ％] than the negative control group maintained in oxaliplatin [(57±3) ％] and the blank group maintained in oxaliplatin [(73t4) ％].Flow cytometry showed that the apoptosis rate of the experimental group maintained in oxaliplatin [(15.07±1.20) ％] was lower than the control group maintained in oxaliplatin [(24.93±0.90) ％] and the blank group maintained in oxaliplatin [(23.71±1.32) ％].Conclusion AP-2α may be related to the sensitivity of colon cancer cells to oxaliplatin.
