槐耳清膏通过氧化应激诱导结直肠癌细胞发生线粒体途径凋亡  

Huaier triggering mitochondria apoptosis in colorectal cancer cells through oxidative stress

作　　者：盛宋昊然 李玉斌 郭家成 孟晓岚 王晨 何淼 李玉强 孙宏治 SHENG Song-hao-ran;LI Yu-bin;GUO Jia-cheng;MENG Xiao-lan;WANG Chen;HE Miao;LI Yu-qiang;SUN Hong-zhi(First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121001,China;Liaoning Provincial Key Laboratory of Clinical Oncology Metabonomics,Jinzhou 121001,China;Clinical Biological Sample Center,First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121001,China)

机构地区：[1]锦州医科大学附属第一医院,辽宁锦州121001 [2]辽宁省肿瘤临床代谢组学重点实验室,辽宁锦州121001 [3]锦州医科大学附属第一医院,临床生物样本中心,辽宁锦州121001

出　　处：《中国中药杂志》2024年第12期3320-3329,共10页China Journal of Chinese Materia Medica

基　　金：辽宁省科学技术计划项目(2021020385-JH2/103);辽宁省教育厅科学技术研究项目(JYTZD2020005)。

摘　　要：该研究探讨了槐耳清膏通过氧化应激诱导结直肠癌线粒体途径凋亡的具体机制。将HCT116与SW480细胞使用槐耳清膏处理后,通过CCK-8、划痕实验分别检测细胞增殖、迁移能力。Annexin-PE染色检测凋亡细胞。DCFH-DA染色与丙二醛(malondialdehyde, MDA)、谷胱甘肽(glutathione, GSH)检测评估细胞氧化应激损伤水平。MitoSOX、JC-1探针分别靶向线粒体活性氧(mitochondria reactive oxygen species, mtROS)与线粒体膜电位(mitochondria membrane potential, MMP)以检测细胞线粒体损伤。Western blot(WB)测定线粒体途径凋亡蛋白与PTEN诱导的推定蛋白激酶1(PTEN-induced putative kinase 1,PINK1)、E3泛素蛋白连接酶Parkin蛋白的表达量。实验表明,在不同浓度槐耳清膏作用下,HCT116、SW480细胞的增殖与迁移能力都受到了抑制,且其可以激活线粒体途径凋亡。与对照组相比,槐耳处理组中结直肠癌细胞的活性氧(reactive oxygen species, ROS)水平升高,MDA上升且GSH下降,提示其发生氧化应激损伤。槐耳处理组的结直肠癌细胞内mtROS上升,MMP下降,表明线粒体损伤。WB实验结果表明,槐耳清膏抑制PINK1/Parkin通路,阻碍了受损线粒体的清除并促进线粒体途径凋亡。该研究表明,槐耳清膏对结直肠癌细胞造成氧化应激与线粒体损伤,抑制PINK1/Parkin通路并促进结直肠癌细胞线粒体途径凋亡。This study investigates the specific mechanisms of Huaier-induced mitochondrial apoptosis in colorectal cancer.HCT116 and SW480 cells were subjected to Huaier treatment.Cell proliferation and migration capabilities were examined through CCK-8 and scratch experiments,respectively.Apoptotic cells were clarified with Annexin-PE staining.DCFH-DA staining,malondialdehyde(MDA),and glutathione(GSH)were used to evaluate the oxidative stress damage level of cells.MitoSOX and JC-1 probes were used to selectively target mitochondria reactive oxygen species(mtROS)and mitochondria membrane potential(MMP)for the evaluation of mitochondria damage.Western blot(WB)experiment was performed to determine apoptosis proteins and PINK1/Parkin pathway.Experiments reveal that in different concentrations of Huaier treatment,the proliferation and migration capabilities of HCT116 and SW480 cells were both restrained.Additionally,mitochondrial apoptosis was activated.Compared with the control group,excessive ROS in colorectal cancer cells was generated in the Huaier group,while MDA increased,and GSH decreased,indicating oxidative stress damage.mtROS increased,and MMP decreased in colorectal cancer cells treated with Huaier,indicating mitochondrial damage.WB result revealed that Huaier suppressed the PINK1/Parkin pathway,hindered the clearance of impaired mitochondria,and subsequently facilitated apoptosis.In conclusion,Huaier impairs colorectal cancer cells through oxidative stress and mitochondria damage.Furthermore,it suppressed the PINK1/Parkin pathway,promoting mitochondria apoptosis in colorectal cancer cells.

关 键 词：槐耳 氧化应激 结直肠癌 线粒体 凋亡 

分 类 号：R285[医药卫生—中药学]