三氧化二砷通过Akt信号通路抑制HGC-27细胞的增殖并促进其凋亡

Arsenic trioxide inhibits HGC-27 cell proliferation and promotes apoptosis through Akt signaling pathway

目的探讨三氧化二砷对HGC-27细胞增殖、凋亡的影响及其作用机制。方法用1μmol/L、2μmol/L、4μmol/L、8μmol/L、16μmol/L的三氧化二砷作用于胃癌细胞HGC-27、MKM28、SGC7901,同时以只加入二甲基亚砜(DMSO)溶液的细胞作为对照组,噻唑蓝(MTT)检测细胞增殖情况,筛选出受到抑制作用最大的细胞继续研究,并计算三氧化二砷半数抑制浓度。用三氧化二砷半数抑制浓度作用于人胃癌细胞,流式细胞术检测细胞凋亡情况,Western blotting检测细胞中活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、蛋白激酶B(Akt)、磷酸化蛋白激酶B(p-Akt)表达水平。用50 ng/ml的Akt信号通路激活剂胰岛素样生长因子-1(IGF-1)及半数抑制浓度的三氧化二砷联合作用于人胃癌细胞作为联合组,以半数抑制浓度的三氧化二砷作用组作为对照,MTT检测细胞增殖情况,流式细胞术检测细胞凋亡情况,Western blotting检测Cleaved Caspase-3、Akt、p-Akt表达水平。结果 1μmol/L、2μmol/L、4μmol/L、8μmol/L、16μmol/L的三氧化二砷能够抑制胃癌细胞HGC-27、MKM28、SGC7901的生长,抑制作用从强到弱依次为:HGC-27、MKM28、SGC7901,半数抑制浓度依次为:(9.34±1.57)μmol/L、(12.92±1.08)μmol/L、(13.24±1.34)μmol/L,后续实验选用9μmol/L的三氧化二砷作用于HGC-27细胞。9μmol/L的三氧化二砷作用后细胞凋亡率从(6.35±2.14)%提高到(39.32±6.54)%,细胞中Cleaved Caspase-3升高,p-Akt水平降低。联合组细胞较单用三氧化二砷组细胞增殖能力增强,细胞凋亡数量减少,细胞中Cleaved Caspase-3水平降低,p-Akt水平升高。结论三氧化二砷能够抑制胃癌细胞增殖,促进胃癌细胞凋亡,激活Akt信号通路能够部分逆转三氧化二砷对胃癌细胞增殖、凋亡的影响。

Objective To investigate the effect and its mechanism of arsenic trioxide on proliferation and apoptosis of HGC-27 cells. Methods 1 μmol/L, 2 μmol/L, 4 μmol/L, 8 μmol/L, 16 μmol/L of arsenic trioxide were injected into gastric cancer cells HGC-27, MKM28, SGC7901, at the same time, only the DMSO solution was added to the control group. Cell proliferation was detected by MTT. The cells with the greatest influence were screened and the half inhibitory concentration was calculated. Half inhibitory concentration of arsenic trioxide was calculated. Cell apoptosis was detected by flow cytometry. Western blotting was used to detect the levels of Cleaved Caspase-3, Akt and p-Akt in cells. The combination of 50 ng/ml Akt signal pathway activator IGF and half inhibitory concentration of arsenic trioxide on human gastric cancer cells were recorded as a combined group. Arsenic trioxide group with half inhibitory concentra- tion was as control. Cell proliferation was detected by MTT. Apoptosis was detected by flow eytometry. Western blotting was used to detect the levels of Cleaved Caspase-3, Akt, p-Akt. Results 1 μmol/L, 2 μmol/L, 4 μmol/L, 8 μmol/L, 16 μmol/L of arsenic trioxide could inhibit the growth of gastric cancer cells HGC-27, MKM28, SGC7901. The inhibi- tory effects from strong to weak were HGC-27, MKM28, SGC7901. The half inhibition concentrations were （9.34 ± 1.57） μmol/L, （12.92 ± 1.08） μmol/L, （13.24± 1.34） μmol/L. The following experiments were conducted with 9μmol/L arsenic trioxide on HGC-27 cells. The rate of apoptosis was increased from （6.35 ± 2.14） % to （39.32 ±6.54） % by the action of arsenic trioxide at a dose of 9 μmol/L, Cleaved Caspase-3 was increased in cells, p-Akt level was decreased. The proliferation ability of the combined group was higher than that of arsenic trioxide group, cell apoptosis was decreased, the level of Cleaved Caspase-3 was decreased in cells, p-Akt level was increased. Conclusion Arsenic trioxide can inhibit the proliferation of gastric cancer cells and promote the apoptosis of gastric cancer cells. The activation of Akt signaling pathway can partly reverse the effects of arsenic trioxide on proliferation and apoptosis of gastric cancer ceils.