中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2011年
7期
489-493
,共5页
细胞分裂周期蛋白42%乳腺肿瘤%MCF-7细胞%雌激素%多药耐药
細胞分裂週期蛋白42%乳腺腫瘤%MCF-7細胞%雌激素%多藥耐藥
세포분렬주기단백42%유선종류%MCF-7세포%자격소%다약내약
Cell division cycle 42%Breast neoplasms%MCF-7 cells%Estrogens%Multidrug resistance
目的 观察细胞分裂周期蛋白42(Cdc42)在雌激素作用下的变化,探讨细胞内物质运输的改变在雌激素引发的乳腺癌细胞耐药中的意义.方法 分别以100 ng/ml 17β雌二醇(E2)处理MCF-7细胞,以Cdc42的小干扰RNA(siRNA) Stealth Select RNAiTM siRNA转染MCF-7细胞.采用四甲基偶氮唑蓝法检测细胞的药物敏感性,流式细胞术检测细胞内阿霉素(ADM)的蓄积量,实时荧光定量聚合酶链反应检测细胞的Cdc42 mRNA表达,Western blot法检测细胞活化的Cdc42蛋白及总Cdc42蛋白表达量.结果 E2处理后,ADM对MCF-7细胞的半数抑制浓度(IC50)由(0.098±0.011) μg/ml增高到(0.134±0.130)μg/ml(P＜0.05),细胞内ADM的相对含量则由7.253±0.310下降为3.233±0.313(P＜0.05),而Cdc42 mRNA、活化Cdc42蛋白及总Cdc42蛋白的表达量均显著增加(P＜0.05).Cdc42 siRNA转染后,ADM对MCF-7细胞的IC50下降到(0.057±0.017)μg/ml(P＜0.05),细胞内ADM的相对含量增高为11.217±0.521(P＜0.05),而Cdc42 mRNA、活化Cdc42蛋白及总Cdc42蛋白则均有显著下降(P＜0.05).结论 雌激素可以诱导乳腺癌细胞耐药性增强,其机制可能是通过上调Cdc42基因的转录、表达和活化,加速胞内物质运输速度,使得化疗药物无法在细胞内聚集.
目的 觀察細胞分裂週期蛋白42(Cdc42)在雌激素作用下的變化,探討細胞內物質運輸的改變在雌激素引髮的乳腺癌細胞耐藥中的意義.方法 分彆以100 ng/ml 17β雌二醇(E2)處理MCF-7細胞,以Cdc42的小榦擾RNA(siRNA) Stealth Select RNAiTM siRNA轉染MCF-7細胞.採用四甲基偶氮唑藍法檢測細胞的藥物敏感性,流式細胞術檢測細胞內阿黴素(ADM)的蓄積量,實時熒光定量聚閤酶鏈反應檢測細胞的Cdc42 mRNA錶達,Western blot法檢測細胞活化的Cdc42蛋白及總Cdc42蛋白錶達量.結果 E2處理後,ADM對MCF-7細胞的半數抑製濃度(IC50)由(0.098±0.011) μg/ml增高到(0.134±0.130)μg/ml(P＜0.05),細胞內ADM的相對含量則由7.253±0.310下降為3.233±0.313(P＜0.05),而Cdc42 mRNA、活化Cdc42蛋白及總Cdc42蛋白的錶達量均顯著增加(P＜0.05).Cdc42 siRNA轉染後,ADM對MCF-7細胞的IC50下降到(0.057±0.017)μg/ml(P＜0.05),細胞內ADM的相對含量增高為11.217±0.521(P＜0.05),而Cdc42 mRNA、活化Cdc42蛋白及總Cdc42蛋白則均有顯著下降(P＜0.05).結論 雌激素可以誘導乳腺癌細胞耐藥性增彊,其機製可能是通過上調Cdc42基因的轉錄、錶達和活化,加速胞內物質運輸速度,使得化療藥物無法在細胞內聚集.
목적 관찰세포분렬주기단백42(Cdc42)재자격소작용하적변화,탐토세포내물질운수적개변재자격소인발적유선암세포내약중적의의.방법 분별이100 ng/ml 17β자이순(E2)처리MCF-7세포,이Cdc42적소간우RNA(siRNA) Stealth Select RNAiTM siRNA전염MCF-7세포.채용사갑기우담서람법검측세포적약물민감성,류식세포술검측세포내아매소(ADM)적축적량,실시형광정량취합매련반응검측세포적Cdc42 mRNA표체,Western blot법검측세포활화적Cdc42단백급총Cdc42단백표체량.결과 E2처리후,ADM대MCF-7세포적반수억제농도(IC50)유(0.098±0.011) μg/ml증고도(0.134±0.130)μg/ml(P＜0.05),세포내ADM적상대함량칙유7.253±0.310하강위3.233±0.313(P＜0.05),이Cdc42 mRNA、활화Cdc42단백급총Cdc42단백적표체량균현저증가(P＜0.05).Cdc42 siRNA전염후,ADM대MCF-7세포적IC50하강도(0.057±0.017)μg/ml(P＜0.05),세포내ADM적상대함량증고위11.217±0.521(P＜0.05),이Cdc42 mRNA、활화Cdc42단백급총Cdc42단백칙균유현저하강(P＜0.05).결론 자격소가이유도유선암세포내약성증강,기궤제가능시통과상조Cdc42기인적전록、표체화활화,가속포내물질운수속도,사득화료약물무법재세포내취집.
Objective To investigate the changes of Cdc42 expression under estrogen stimulation, and to explore the signaling pathway of intracellular material transportation caused by estrogen. Methods MTT was used to test the drug sensitivity of cells. Real-time PCR was used to evaluate the expression of Cdc42 mRNA. The amount of ADM accumulated in MCF-7 cells was detected by flow cytometry. The protein levels of active-Cdc42 and Total-Cdc42 were measured by Western blot. Results IC50 of ADM in MCF-7 cells was increased from (0.098±0.011)μg/ml to (0.134±0.130)μg/ml (P<0.05) after estrogen stimulation. The amount of ADM accumulated in MCF-7 cells was reduced from 7.253±0.310 to 3.233±0.313 (P<0.05). All of Cdc42 mRNA, active-Cdc42 protein and total-Cdc42 protein were increased (P<0.05). After the treatment with siRNA, the IC50 of ADM in siRNA group was decreased to (0.057±0.017)μg/ml (P<0.05) compared with that in the control group. The amount of accumulated ADM was significantly increased in the siRNA group, and all the expression levels of Cdc42 mRNA, active-Cdc42 protein and total-Cdc42 protein were decreased in the siRNA group (P<0.05). Conclusions Estrogen enhances the drug resistance in breast cancer cells. The mechanism of this effect may be via the enhancing Cdc42 expression and decreasing the accumulation of chemotherapeutic drugs in the cancer cells.
