食道通结方对食管癌细胞株TE-1增殖和凋亡的影响

目的观察具有理气化痰功效的食道通结方对食管癌细胞株增殖和凋亡的影响。方法利用SD大鼠制备食道通结方含药血清,以食管癌TE-1细胞株为观察对象,用MTT法筛选合适的含药血清浓度;再用含药血清、顺铂和含药血清+顺铂分别作用于TE-1细胞株。采用MTT检测各组肿瘤细胞的生长情况,流式细胞仪检测各组细胞周期及细胞凋亡情况,Western印迹法检测各组凋亡相关蛋白(p-Akt、Akt、P53、Bcl-2、Bax、Caspase3)的表达。结果①确定20%的含药血清为最佳含药血清浓度。②含药血清组、顺铂组和含药血清+顺铂组细胞增殖抑制率均随干预时间的延长(0 h,12 h,24 h,48 h)而增加,48 h时的细胞增殖抑制率最高,分别为30.9%、35.4%和50.6%,组间差异有统计学意义(P0.01)。③与对照组比较,顺铂组和含药血清+顺铂组G1期细胞比例增高(P0.01),含药血清组和含药血清+顺铂组G2期细胞比例降低(P0.05);含药血清+顺铂组G1期细胞比例高于顺铂组和含药血清组,差异有统计学意义(P0.01)。④12 h、24 h组内比较,各组细胞凋亡指数差异均有统计学意义(P0.01);与对照组各观察时点比较,顺铂组、含药血清组和含药血清+顺铂组细胞凋亡指数均明显增加(P0.01);顺铂组、含药血清+顺铂组各观察时点细胞凋亡指数均高于含药血清组(P0.01),含药血清+顺铂组各观察时点细胞凋亡指数高于顺铂组(P0.01)。⑤与对照组比较,顺铂组、含药血清组和含药血清+顺铂组中p-Akt、Bcl-2表达明显下降(P0.01),P53、Bax、Caspase3表达显著增加(P0.01)。结论食道通结方含药血清对食管癌TE-1细胞株的增殖和生长有一定的抑制作用,能够促进细胞凋亡,与顺铂联合后具有较为明显的协同作用。

Effect of ＂ Shidao Tongjie Decoction＂ on proliferation and apoptosis of esophageal cancer cell lines TE-1

Objective To investigate the effect of ＂ Shidao Tongjie Decoction＂ （SDTJ） on proliferation and apoptosis of esophageal cancer cell lines TE-1. Methods SD rats were established by serum containing SDTJ. The esophageal cancer cell lines TE-1 was used as the object. The appropriate concentration of drug serum was screened by MTT assay; Then serum, serum combined with cisplatin and cisplatin were applied to TE-1 cell lines. The growth of cancer cells were detected by MTY assay, and the cell cycle, apoptosis were detected by FCM. The expressions of apoptosis-related proteins （p-Akt, Akt, P53, Bcl-2, Bax, and Caspase3） were detected by Western blotting. Resets ① The optimum concentration of the drug serum was 20%. ② MTT results showed that inhibition rates of serum group, cisplatin group and cisplatin plus serum group were fortified as the time goes by, and at 48h were 30.9%, 35.4% and 50.6%. There was a significant difference among the groups （ P 〈 0.01 ）. ③ Compared with the control group, the proportion in G1 phase of the cisplatin group and cisplatin plus serum group were significantly higher （P 〈0.01 ）, and the proportion in G2 phase of the serum group and cisplatin plus serum group were reduced （ P 〈 0.05 ） ; The proportion in Gt phase of the cisplatin plus serum group was higher than those of the cisplatin group and serum group, with significant difference between them （P 〈0.01 ）. ④ FCM showed that there was a significant differences in apoptosis index of each intervention group at 12h and 24h （P 〈 0.01 ） ; Compared with control group at different time-points, the apoptotic indexes of the cisplatin group, serum group and cisplatin plus serum group was markedly increased （ P 〈0.01 ） ; the apoptotic indexes of the cisplatin group, cisplatin plus serum group were higher than that of the drug serum group （ P 〈 0.01 ）, the apoptotic index at different time-points of the cisplatin plus serum group higher than that of cisplatin group （ P 〈0.01 ）. ⑤ Compared with the control group, the expressions of p-Akt and Bcl-2 in serum group, cisplatin group and cisplatin plus serum group were reduced （ P 〈 0.01 ）, the expressions of P53, Bax and Caspase3 were strengthened （P 〈0.01 ）. Conclusion Serum containing SDTJ might have the effect of inhibiting proliferation and promoting apoptosis on esophageal TE-1 cell line. And there shows the obvious synergies when it combined with cisplatin.