小檗碱增强宫颈癌细胞放射敏感性的研究

Effect of berberine on radiosensitivity of cervical cancer cells

目的 探讨小檗碱对宫颈癌细胞凋亡及放射敏感性的影响。方法 用5、10、15、20 μmol/L的小檗碱作用于宫颈癌细胞Siha、HeLa、Caski记为给药组，同时以只加入二甲基亚砜（DMSO）的细胞为对照组，CCK-8法检测48 h细胞存活率，计算小檗碱半数抑制浓度。用半数抑制浓度的小檗碱作用于宫颈癌Siha细胞48 h，流式细胞术检测细胞凋亡率和细胞周期，Western blot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）、细胞周期素B1（Cyclin Bl）、细胞周期蛋白依赖性激酶1（CDK1）、信号转导和转录因子（STAT）、磷酸化的STAT3（p-STAT3）表达水平。含有半数抑制浓度的小檗碱培养液培养宫颈癌Siha细胞24 h后，用0、2、4、6、8 Gy X射线照射细胞，细胞克隆形成实验检测放射敏感性。结果 小檗碱能够抑制宫颈癌细胞的生长，抑制作用从大到小为：Siha、Caski、HeLa，半数抑制浓度依次为（16.84±3.52）、（23.54±8.67）、（21.86±6.35）μmol/L。17 μmol/L的小檗碱能够促进宫颈癌细胞Siha凋亡（t=56.847，P〈0.01），将细胞周期阻滞在G2/M期（t=47.251，P〈0.01），并抑制宫颈癌细胞中Cyclin B1、CDK1、p-STAT3表达，促进Cleaved Caspase-3表达，对宫颈癌细胞中STAT3表达没有影响。17 μmol/L的小檗碱能够降低宫颈癌细胞的存活分数，放射增敏比为1.550。结论 小檗碱能够抑制宫颈癌细胞增殖，促进凋亡，阻滞细胞周期，增加宫颈癌细胞的放射敏感性。

Objective To investigate the effect of berberine on the radiosensitivity of cervical cancer cells. Methods 5, 10, 15, 20 μmol/L of berberine were used to treat cervical cancer cell lines of Siha, HeLa, Caski. DMSO was applied as control of drug treatment. Cell proliferation was detected by the CCK-8 method, and then the half inhibitory concentration of berberine was calculated. Cell apoptosis and cell cycle distribution were detected by flow cytometry. Protein expressions of Cleaved Caspase-3, Cyclin B1, CDK1, STAT3 and p-STAT3 were detected by Western blot. Cervical cancer cells of Siha were treated by berberine with a half inhibitory concentration for 24 h and then irradiated with 0, 2, 4, 6, 8 Gy of X-rays. Cell clone assay was used to detect cell survival. Results Berberine could inhibit the growth of cervical cancer cells with a half inhibition concentration of（16.84±3.52）,（23.54±8.67）,（21.86±6.35）μmol/L for Siha, Caski, and HeLa cells, respectively. The berberine at 17 μmol/L could induce apoptosis（t=56.847,P〈0.01）and G2/M phase arrest（t=47.251,P〈0.01）in Siha cells, which also inhibited the expressions of Cyclin B1, CDK1 and p-STAT3 and promoted the expression of cleaved Caspase-3, but did not influence the expression of STAT3 in cervical cancer cells. Treatment of cells with 17 μmol/L berberine increased the radiosensitivity of cervical cancer cells with a sensitivity enhancement ratio of 1.55. Conclusions Berberine can inhibit cell proliferation, promote apoptosis, block cell cycle, and increase radiosensitivity of cervical cancer cells.