Zhang Xin, Zhou Xuebing, Gao Ming, Lyu You, Wang Ying, Yang Chunyu, Piao Yingshi, Ren Xiangshan
Cancer Research Center/Key Laboratory of the Science and Technology Department of Jilin Province, Yanbian University, Medical College of Yanbian University, Yanji 133002, China.
Medical College of Yanbian University, Medical College of Yanbian University, Yanji 133002, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2022 Jun;38(6):513-521.
Objective To explore the inhibitory effect of cordycepin on the proliferation and migration of gastric cancer cells and its molecular mechanism. Methods MGC-803 cells were treated with 0, 25, 50, 100 μmol/L of cordycepin and HGC-27 cells with 0, 5, 25, 50 μmol/L of cordycepin for 48 hours. The proliferation ability of MGC-803 and HGC-27 cells was detected by MTT assay and EdU assay; the colony formation ability of cells was detected by colony formation assay; both wound healing assay and cell migration assay were applied to detect the cell migration ability of MGC-803 and HGC-27 cells; the chromatin agglutination was detected by Hoechst 33342 staining; the apoptosis of gastric cancer cells was detected by annexin V-FITC/PI double labeling combined with flow cytometry; Western blot was used to measure the protein expression levels of lipid metabolism-related proteins including sterol regulatory element binding transcription factor 1 (SREBF1), fatty acid synthase (FASN), and acetyl coA carboxylase 1 (ACC1), epithelial-mesenchymal transition (EMT)-related proteins including E-cadherin, vimentin, Snail, Slug, matrix metalloproteinase 2 (MMP2), MMP9, AMPK, and phosphorylated AMPK (p-AMPK), MAPK signaling pathway-related proteins including JNK, phosphorylated JNK (p-JNK), p38 MAPK, and p-p38 MAPK, and apoptosis-related proteins including cleaved caspase-9 (c-caspase-9), c-caspase-3, and cleaved poly (ADP-ribose) polymerase (c-PARP). Results Cordycepin significantly inhibited the proliferation, colony formation, and migration of gastric cancer cells. After cordycepin treatment, the karyopycnosis, karyorrhexis, and apoptosis rate of cancer cells increased, and the expressions of apoptosis-related proteins c-caspase-3, c-caspase-9, and c-PARP increased. The expression of E-cadherin increased, while the expressions of vimentin, Snail, Slug, SREBF1, FASN, ACC1, MMP2, MMP9 significantly decreased; the phosphorylation levels of AMPK, JNK and P38 proteins significantly increased. Conclusion Cordycepin inhibits the proliferation and migration of gastric cancer cells by suppressing the lipid metabolism and EMT process via activating AMPK and MAPK signaling pathway.
目的 探讨虫草素对胃癌细胞增殖和迁移的抑制作用及其分子机制。方法 用0、25、50、100 μmol/L虫草素处理MGC-803细胞，用0、5、25、50 μmol/L虫草素处理HGC-27细胞48小时。采用MTT法和EdU法检测MGC-803和HGC-27细胞的增殖能力；采用集落形成试验检测细胞的集落形成能力；采用划痕愈合试验和细胞迁移试验检测MGC-803和HGC-27细胞的迁移能力；采用Hoechst 33342染色检测染色质凝集；采用膜联蛋白V-FITC/PI双标记结合流式细胞术检测胃癌细胞凋亡；采用蛋白质免疫印迹法检测脂质代谢相关蛋白包括固醇调节元件结合转录因子1（SREBF1）、脂肪酸合酶（FASN）和乙酰辅酶A羧化酶1（ACC1）、上皮-间质转化（EMT）相关蛋白包括E-钙黏蛋白、波形蛋白、Snail、Slug、基质金属蛋白酶2（MMP2）、MMP9、AMPK和磷酸化AMPK（p-AMPK）、丝裂原活化蛋白激酶（MAPK）信号通路相关蛋白包括JNK、磷酸化JNK（p-JNK）、p38 MAPK和磷酸化p38 MAPK（p-p38 MAPK）以及凋亡相关蛋白包括裂解的半胱天冬酶-9（c-caspase-9）、c-caspase-3和裂解的聚（ADP-核糖）聚合酶（c-PARP）的蛋白表达水平。结果 虫草素显著抑制胃癌细胞的增殖、集落形成和迁移。虫草素处理后，癌细胞的核固缩、核碎裂和凋亡率增加，凋亡相关蛋白c-caspase-3、c-caspase-9和c-PARP的表达增加。E-钙黏蛋白的表达增加，而波形蛋白、Snail、Slug、SREBF1、FASN、ACC1、MMP2、MMP9的表达显著降低；AMPK、JNK和P38蛋白的磷酸化水平显著增加。结论 虫草素通过激活AMPK和MAPK信号通路抑制脂质代谢和EMT过程，从而抑制胃癌细胞的增殖和迁移。
