摘要
目的 检测miR-219a-5p在胃癌及其癌旁组织中的表达并分析其启动子区域甲基化水平,探讨其对胃癌细胞系生物学功能的影响。方法 利用实时荧光定量PCR (Real-time quantitative polymerase chain reaction, RT-qPCR)和甲基化特异性PCR (methylation specific PCR,MSP)方法检测胃癌及其癌旁组织中miR-219a-5p的表达及甲基化水平,通过细胞增殖实验和流式细胞仪检测过表达miR-219a-5p后对胃癌细胞增殖和凋亡的影响;体外细胞迁移和侵袭实验检测肿瘤细胞迁移和侵袭能力。结果 miR-219a-5p的表达水平在胃癌患者的癌组织中呈现普遍下调,miR-219a-5p在癌组织中的相对表达量(0.769±0.95)明显低于癌旁组织表达(2.114±5.10),差异具有统计学意义(P<0.05);且miR-219a-5p表达下调组的DNA甲基化水平比表达上调组明显升高;过表达miR-219a-5p的胃癌细胞系MGC-803细胞增殖能力和迁移侵袭能力明显受到抑制(P<0.05),过表达组凋亡比例明显增高(P<0.05),凋亡相关蛋白caspase-3的表达水平明显升高(P<0.05)。结论 miR-219a-5p在胃癌组织中受甲基化调控普遍低表达,且其具有抑制细胞增殖和迁移的能力,促进细胞凋亡等类似抑癌基因的作用,有望成为胃癌新的诊疗靶点。
Objective To assess miR-219 a-5 p expression and its promoter methylation level in gastric cancer and adjacent tissues, and to explore its effect on the biological functions of gastric cancer cell lines. Methods The expression and methylation level of miR-219 a-5 p in gastric cancer and adjacent tissues were detected by qPCR and MSP. Proliferation and apoptosis of gastric cancer cells were detected by Cell Counting Kit-8 assays and flow cytometry after overexpression of miR-219 a-5 p. Results MiR-219 a-5 p expression was generally downregulated in gastric cancer tissues. The relative expression of miR-219 a-5 p in cancer tissues(0.769 ± 0.95) was significantly lower than that in adjacent tissues(2.114 ± 5.10)(P<0.05). The DNA methylation level in the miR-219 a-5 p-downregulated group was significantly higher than that in the miR-219 a-5 p-upregulated group. Proliferation and invasion of MGC-803 cells overexpressing miR-219 a-5 p were inhibited significantly(P<0.05), apoptosis in the overexpression group was increased significantly(P<0.05), and expression of apoptosis-related protein caspase-3 was significantly higher(P<0.05). Conclusions MiR-219 a-5 p is generally underexpressed by methylation in gastric cancer tissue, and inhibits cell proliferation and migration, promotes apoptosis, and play a role similar to tumor suppressor genes. It is expected to become a new target for diagnosis and treatment of gastric cancer.
作者
罗敏
章帆
王利民
李红霞
卿克勤
LUO Min;ZHANG Fan;WANG Limin;LI Hongxia;QING Keqin(Department of Laboratory Medicine,Chengdu First People’s Hospital,Chengdu 610000,China)
出处
《中国比较医学杂志》
CAS
北大核心
2022年第7期66-73,共8页
Chinese Journal of Comparative Medicine
基金
2020年四川省卫生健康委员会科研课题(20PJ188)。
