健脾清热活血方对miR-222基因沉默转染结肠癌HCT-116细胞的生物学作用及MAPK10、PANK3表达的影响  

Biological effect of Jianpi Qingre Huoxue Formula and the expression of MAPK10 and PANK3 in colon cancer HCT-116 cells transfected by miR-222 gene silencing

作　　者：赖冬萍 卢鑫 张涛[1] 陈小芬 钟婵 黄李冰雪 黄晓燕[3] 李锦颜 朱梓铭[1] LAI Dongping;LU Xin;ZHANG Tao;CHEN Xiaofen;ZHONG Chan;HUANG Libingxue;HUANG Xiaoyan;LI Jinyan;ZHU Ziming(Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530000,China;Guangxi University ofChinese Medicine,Nanning 530000,China;The First Affiliated Hospital of Guangxi University ofChinese Medicine,Nanning 530000,China)

机构地区：[1]广西中医药大学附属瑞康医院,南宁530000 [2]广西中医药大学,南宁530000 [3]广西中医药大学第一附属医院,南宁530000

出　　处：《中华中医药杂志》2025年第1期359-364,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81260536);广西自然科学基金项目(No.2018GXNSFAA281042);广西中医药大学一流学科项目(No.2018XK082);广西研究生教育创新计划(No.YCSY2020050,No.YCXJ2021109)。

摘　　要：目的:研究健脾清热活血方含药血清对稳定沉默miR-222结肠癌HCT-116细胞株生物学作用及丝裂原活化蛋白激酶10(MAPK10)、泛酸激酶3(PANK3)表达变化。方法:采用慢病毒介导的RNA干扰技术沉默miR-222基因,培养细胞分为:空白对照组(HCT-116)、阴性对照组、阳性对照组(sh-miR-222)和各剂量治疗组(sh-miR-222+低、中、高剂量中药)。细胞增殖检测试剂比色法检测细胞增殖,流式细胞术检测细胞凋亡及周期,Transwell检测细胞迁移;透射电镜观察细胞自噬。RT-PCR及Western Blot检测MAPK10、PANK3 mRNA及蛋白表达。结果:与空白对照组和阳性对照组比较,治疗组抑制HCT-116细胞增殖和生长活力的效应明显加强(P<0.05),细胞总凋亡率显著升高,细胞迁移数显著下降(P<0.01)。治疗组的细胞胞浆内可见明显增多的自噬体,与阳性对照组比较,高剂量治疗组MAPK10、PANK3 mRNA表达量和低剂量治疗组MAPK10、PANK3蛋白表达量显著升高(P<0.01)。结论:健脾清热活血方联合miR-222基因沉默可增强诱导结肠癌HCT-116细胞凋亡和自噬,减少细胞增殖和迁移,MAPK10、PANK3表达显著升高。Objective:To study the biological effect of Jianpi Qingre Huoxue Formula on stable silencing miR-222 colon cancer HCT-116 cell line and the changes of mitogen-activated protein kinase10(MAPK10) and pantothenate kinase 3(PANK3)expression.Methods:Lentivirus-mediated RNA interference technique was used to silence miR-222 gene.The cultured cells were divided into blank control group(HCT-116),negative control group,positive control group(sh-miR-222) and treatment group(sh-miR-222+low/medium/high dose of traditional Chinese medicine).MTS assay was used for detecting cell proliferation,flow cytometry for detecting apoptosis and cycle,and transwell for detecting cell death and cycle.Transwell was used for detecting cell migration;transmission electron microscopy was sued to observe cell autophagy.RT-PCR and Western Blot was used to detect the m RNA and protein expressions of MAPK10 and PANK3.Results:Compared with the blank control group and the positive control group,the inhibitory effect on the proliferation and growth vitality of HCT-116 cells in the treatment group was significantly enhanced(P<0.05),the total apoptosis rate was significantly increased,and the number of cell migration decreased significantly(P<0.01).Autophagosomes were significantly increased in the cytoplasm of cells in the treatment group.Compared with the positive control group,the mRNA expression in high dose treatment group and protein expression of MAPK10 and PANK3 in low dose treatment group were significantly up-regulated(P<0.01).Conclusion:Jianpi Qingre Huoxue Formula combined with miR-222 gene silencing can enhance the induction of apoptosis and autophagy of colon cancer HCT-116 cells,reduce cell proliferation and migration,and significantly increase the expression of MAPK10 and PANK3.

关 键 词：健脾清热活血方 miR-222基因 结肠癌细胞 丝裂原活化蛋白激酶10 泛酸激酶3 

分 类 号：R285[医药卫生—中药学]