摘要
目的:探讨苦瓜皂苷对人恶性黑素瘤SKMEL-2细胞生长及JAK/STAT5通路的影响。方法:采用1、2.5、5、7.5、10mg/mL浓度的苦瓜皂苷处理体外培养的人恶性黑素瘤SKMEL-2细胞,以CCK-8法分别在24、48、72h后检测细胞增殖情况,筛选合适药物作用浓度和时间。体外培养的SKMEL-2细胞,随机分为三组:对照组(control)、苦瓜皂苷组及顺铂组,药物处理后,CCK-8检测细胞增殖情况,测定细胞存活率;流式细胞技术检测细胞凋亡情况,测定细胞凋亡率;采用免疫印记(WB)法检测caspase3、Bcl-2、Bax、JAK、p-JAK、STAT5、p-STAT5的蛋白表达水平。结果:药物作用48、72h其抑制作用比24h高,而药物作用48、72h其抑制作用基本相同;浓度为1~5、7.5、10mg/mL的苦瓜皂苷都能抑制SKMEL-2增殖,其抑制作用随浓度升高而增强,但在药物作用48、72h后,浓度达到5mg/mL时抑制作用趋于稳定。选择5mg/mL浓度的苦瓜皂苷处理SKMEL-2细胞48h用于后续实验;药物处理后,与control组比较,苦瓜皂苷组及顺铂组细胞相对存活率、p-JAK、p-STAT5、Bcl-2表达明显降低(P<0.05),细胞凋亡率、caspase3、Bax表达明显升高(P<0.05),JAK、STAT5表达无明显变化,差异无统计学意义(P>0.05);苦瓜皂苷组与顺铂组相比,各指标无明显变化,差异无统计学意义(P>0.05)。结论:苦瓜皂苷对人恶性黑素瘤SKMEL-2细胞的生长有抑制作用、对其凋亡有促进作用,下调JAK/STAT5通路活性,上调凋亡蛋白caspase3、Bax表达,抑制抗凋亡蛋白Bcl-2表达可能是其作用机制。
Objective:To investigate the effects of balsam pear saponins on the growth of human malignant melanoma SKMEL-2 cells and JAK/STAT5 pathway.Methods:Cultured human malignant melanoma SKMEL-2 cells were treated with balsam pear saponins at concentrations of 1,2.5,5,7.5 and 10 mg/mL.The proliferation of SKMEL-2 cells was detected by CCK-8 method after 24,48 and 72 hours,respectively,and the appropriate concentration and time of drug action were screened.SKMEL-2 cells cultured in vitro were randomly divided into three groups:control group,balsam pear saponin group and cisplatin group.After drug treatment,CCK-8 was used to detect cell proliferation,cell viability was measured,and cell apoptosis was detected by flow cytometry,the apoptotic rate was measured.The expression levels of caspase 3,Bcl-2,Bax,JAK,p-JAK,STAT5 and p-STAT5 were detected by immunoblotting(WB).Results:The inhibitory effect of the drug was higher at 48 hours and 72 hours than that at 24 hours,while the inhibitory effect was basically the same at 48 hours and 72 hours.The proliferation of SKMEL-2 was inhibited by balsam pear saponins at concentrations of 1-5,7.5 and 10 mg/mL,and the inhibition increased with the increase of concentration,but after 48 and 72 hours,the inhibitory effect tended to be stable when the concentration reached 5 mg/mL.SKMEL-2 cells were treated with balsam pear saponins at a concentration of 5 mg/mL for 48 hours.Compared with control group,the relative survival rate,expressions of p-JAK,p-STAT5 and Bcl-2 in balsam pear saponin group and cisplatin group decreased significantly(P<0.05),apoptotic rate,caspase 3 and Bax expression were significantly increased(P<0.05),there was no significant difference in JAK and STAT5 expressions(P>0.05).There was no significant change in each index between balsam pear saponin group and cisplatin group(P>0.05).Conclusion:Momordica charantia saponins can inhibit the growth of SKMEL-2 cells,promote their apoptosis.It may be the mechanism that momordica charantia saponins can down-regulate the activity of JAK/STAT5 pathway,up-regulate the expressions of caspase 3 and Bax,and inhibit the expression of Bcl-2.
作者
黄翠敏
叶秀群
骆月姬
HUANG Cuimin;YE Xiuqun;LUO Yueji(The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangdong Guangzhou 510405, China)
出处
《河北医学》
CAS
2020年第4期563-568,共6页
Hebei Medicine
