摘要
目的研究奥沙利铂联合柔红霉素对结肠癌细胞凋亡的影响及机制。方法分别用不同浓度的奥沙利铂和柔红霉素处理结肠癌细胞HT-29,MTT法检测细胞增殖并计算半数抑制浓度。分别用半数抑制浓度的奥沙利铂、柔红霉素及二者联合处理HT-29细胞,MTT法测定细胞增殖;平板克隆实验检测细胞克隆形成;流式细胞计量术检测细胞凋亡;Western blot检测细胞中活化的caspase-3(cleaved caspase-3)、第10号染色体同源缺失性磷酸酶-张力蛋白(PTEN)、磷酸化Akt(p-Akt)蛋白表达。结果不同浓度的奥沙利铂和柔红霉素均可以抑制HT-29细胞增殖(P<0. 05),其半数抑制浓度为:柔红霉素约0. 23μmol/L,奥沙利铂约40 mg/L。柔红霉素、奥沙利铂和奥沙利铂联合柔红霉素处理后的HT-29细胞增殖能力、克隆形成能力降低;细胞凋亡率升高;细胞中cleaved caspase-3蛋白表达升高;细胞中p-Akt蛋白表达降低;PTEN蛋白表达升高。奥沙利铂联合柔红霉素作用高于单纯柔红霉素或奥沙利铂处理(P<0. 05)。结论奥沙利铂联合柔红霉素能够诱导结肠癌细胞HT-29凋亡,其作用机制可能与PTEN/Akt信号调控有关。
Objective To study the mechanism of oxaliplatin combined with daunorubicin on the apoptosis of colon cancer cells. Methods Colon cancer cells HT-29 were incubated with different concentrations of oxaliplatin and daunorubicin respectively,MTT assay was used to detect cell proliferation and calculate the median inhibitory concentration.HT-29 cells were treated with half inhibitory concentration of oxaliplatin,daunorubicin and by two drugs combined. MTT assay cell proliferation ability,the colony forming ability of cells was detected by plate cloning test,cell apoptosis was detected by flow cytometry,Western blot was used to detect the expression of cleaved caspase-3,PTEN and p-Akt in cells. Results Different concentrations of oxaliplatin and daunorubicin inhibited the proliferation of HT-29 cells,the half inhibitory concentration of daunorubicin was about 0. 23 μmol/L and oxaliplatin was about40 mg/L. Daunorubicin,oxaliplatin or oxaliplatin plus daunorubicin were used to treat HT-29 cells,the proliferation of cells decreased,the ability of clone formation also reduced,the rate of apoptosis increased,the expression of cleaved caspase-3 protein increased in cells,the expression of p-Akt protein in cells decreased,the expression of PTEN protein elevated( P < 0. 05). The effect of oxaliplatin combined with daunorubicin was higher than that of daunorubicin or oxaliplatin alone( P<0. 05). Conclusions Oxaliplatin combined with daunorubicin can induce apoptosis in colon cancer cells,its mechanism may be related to the regulation of PTEN/Akt signaling.
作者
田春阳
刘晓政
范军朝
TIAN Chun-yang;LIU Xiao-zheng;FAN Jun-chao(Department of Gastroenterology,Nanyang Central Hospital,Nanyang 473000;Department of Anesthesiology,the First Affiliated Hospital of Henan University,Zhengzhou 475000,China)
出处
《基础医学与临床》
CSCD
2020年第3期310-314,共5页
Basic and Clinical Medicine
基金
河南省教育厅科研基金(16B320003)
