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Received:March 08, 2023 Published Online:July 19, 2023
Received:March 08, 2023 Published Online:July 19, 2023
中文摘要: 目的 探讨miR-495-3p在宫颈癌患者癌组织中的表达及其对人宫颈癌细胞HeLa细胞凋亡和侵袭的影响。方法 组织学实验： 选取2020年12月至2021年12月河北中石油中心医院和石家庄市第四医院归档的资料齐全完整的47例宫颈癌患者宫颈组织及其癌旁组织手术标本，通过实时荧光定量PCR对癌组织和癌旁组织miR-495-3p表达水平进行检测。细胞学实验： 将人宫颈癌细胞HeLa分为3组，对照组（不转染）、NC组（转染NC质粒）和miR-495-3p组（转染miR-495-3p质粒），划痕实验和细胞迁移实验(Transwell）检测各组细胞迁移和侵袭能力，流式细胞仪检测各组细胞凋亡水平及细胞周期；Western Blot检测G1/S-特异性周期蛋白D1（Cyclin D1）的表达水平。结果 组织学实验表明，与癌旁组织相比，癌组织中miR-495-3p低表达（0.396±0.012 vs 1.893±〖JP〗0.215, t=47.663, P＜0.01）。细胞学实验表明，与对照组相比，miR-495-3p组细胞迁移能力和侵袭能力显著下降（P＜0.01）；细胞存活比例显著降低，早期凋亡和晚期凋亡比例显著增加（P＜0.05）；G0/G1时期细胞比例显著增加，S期和G2期细胞比例显著降低(P＜0.05)；Western Blot实验表明miR-495-3p组CyclinD1蛋白表达水平较其他两组显著降低（P＜0.01），NC组细胞和对照组细胞差异无统计学意义（P>0.05）。结论 miR-495-3p在宫颈癌组织中显著低表达，miR-495-3p可以通过抑制宫颈癌细胞的迁移和侵袭能力，促进细胞凋亡，同时通过抑制CyclinD1的表达干预宫颈癌细胞周期调控。
Abstract:Objective To investigate the expression of miR-495-3p in cervical carcinoma tissue and its effects on apoptosis and invasion of human cervical carcinoma cell line Hela.
Methods The surgical specimens of cervical carcinoma tissue and its adjacent tissue were collected from 47 cervical carcinoma patients with complete data archived in Hebei Petro China Central Hospital and Shijiazhuang Fourth Hospital from December 2020 to December 2021. Quantitative polymerase chain reaction was used to detect the expression levels of miR-495-3p in cancer tissues and adjacent tissues. The HeLa cells were divided into three groups： control group(non-transfected), NC group(transfected with NC plasmid) and miR-495-3p group(transfected with miR-495-3p plasmid). In three groups of Hela cells, scratch and Transwell assay were respectively used to detect the ability of cell migration and invasion, flow cytometry was used to check the apoptosis and cell cycle status,and Western Blot was used to test the protein expression level of G1/s-specific cyclin-D1 (cyclin D1).
Results Histological experiment showed that the expression of miR-495-3p in cancer tissues was significantly lower than that in adjacent tissue(0.396±0.012 vs 1.893±0.215, t=47.663, P＜0.01). Compared with those in control group, the cell migration ability and invasion ability decreased significantly(P＜0.01), the proportion of cell survival decreased significantly, and early and late apoptosis increased significantly in miR-495-3p group(P＜0.05). The proportion of cells increased significantly in G0/G1 phase, and the proportion of cells in S phase and G2 phase decreased significantly(P＜0.05). Western Blot showed that the expression level of CyclinD1 protein in miR-495-3p group decreased significantly compared with that in other two groups(P＜0.01), and there was no significant difference in it between NC group and control group(P>0.05).
Conclusion The expression of miR-495-3p is obviously low in cervical carcinoma tissues. miR-495-3p can promote cell apoptosis by inhibiting the migration and invasion of cervical carcinoma, and regulate the cervical carcinoma cell cycle by inhibiting the expression of CyclinD1.
文章编号： 中图分类号：R737.33 文献标志码：A
基金项目:河北省医学科学研究课题计划（20222694）
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