乙酰紫草素联合奥沙利铂对人结肠癌HT29细胞增殖及凋亡的影响

目的探讨乙酰紫草素联合奥沙利铂对人结肠癌HT29细胞增殖及凋亡的影响。方法将人结肠癌HT29细胞分为空白组、乙酰紫草素组(10μmol/L)、奥沙利铂组(1μmol/L)和联合给药组(10μmol/L乙酰紫草素+1μmol/L奥沙利铂),MTT法检测HT29细胞增殖抑制率,流式细胞术检测HT29细胞凋亡率、qRT-PCR法检测HT29细胞Ki-67、Bcl-2及Bax mRNA表达,Western blot法检测磷酸化磷脂酰肌醇-3-羟激酶(p-PI3K)、磷酸化蛋白激酶B (p-Akt)蛋白表达。结果人结肠癌HT29细胞经乙酰紫草素、奥沙利铂和联合给药处理24、48、72 h后,细胞增殖抑制率均增加,其中联合给药组细胞增殖抑制率最高(P<0. 05)。药物处理HT29细胞48 h后,与空白组比较,乙酰紫草素组、奥沙利铂组和联合给药组凋亡率、Bax mRNA表达增加(P<0. 05),而Ki-67、Bcl-2 mRNA表达及p-PI3K、p-Akt蛋白表达降低(P<0. 05);联合给药组的细胞凋亡率、Bax mRNA表达均高于乙酰紫草素组和奥沙利铂组,Ki-67、Bcl-2 mRNA表达及p-PI3K、p-Akt蛋白表达低于乙酰紫草素组和奥沙利铂组。结论乙酰紫草素和奥沙利铂对人结肠癌HT29细胞均具有抑制增殖及诱导凋亡作用,可能与调控PI3K/Akt信号通路有关,且两者联用时效果更好。

Effects of acetyl shikonin combined with oxaliplatin on the proliferation and apoptosis of human HT29 colon cancer cells

AIM To investigate the effect of acetyl shikonin combined with oxaliplatin on the proliferation and apoptosis of human HT29 colon cancer cells. METHODS Human HT29 colon cancer cells were divided into blank group,acetyl shikonin group(10 μmol/L),oxaliplatin group(1 μmol/L),and combinative administration group(10 μmol/L acetyl shikonin+1 μmol/L oxaliplatin). HT29 cells were subjected to the determination of proliferation inhibition rate by MTT and the apoptosis rate by flow cytometry,the identifications of expression of Ki-67,Bcl-2 and Bax mRNA by qRT-PCR,and the expression of phosphorylated phosphatidylinositol-3-hydroxykinase( pPI3 K) and phosphorylated protein kinase B( p-Akt) by Western blot. RESULTS Human HT29 colon cancer cells treated with acetyl shikonin,or oxaliplatin,or combinative administration for 24,48,72 h shared increased cell proliferation inhibition rates,and the combinative administration group was observed with the highest cell proliferation inhibition rate( P<0. 05). Compared with the blank group,all of the groups exposed to 48 hours of drug treatment had increased apoptosis rate and Bax mRNA expression in HT29 cells( P<0. 05),reduced Ki-67,Bcl-2 mRNA expression and p-PI3 K,p-Akt protein expression( P<0. 05). The combination administration group demonstrated higher apoptosis rate and Bax mRNA expression,lower Ki-67,Bcl-2 mRNA expression and p-PI3 K,p-Akt protein expression than the acetyl shikonin group and oxaliplatin group. CONCLUSION Both acetyl shikonin and oxaliplatin are antiproliferative and cytotoxic,and their combinative use may produce a synergetic effect on human HT29 colon cancer cells becausee of their effect on PI3 K/Akt signaling pathway.