miR-214靶向Lnc-PVT1抑制乳腺癌细胞对PD1抑制剂敏感性

目的： 探讨MiR-214靶向Lnc-PVT1抑制乳腺癌细胞对PD1抑制剂敏感性的分子机制。方法： 通过实时荧光定量PCR实验（quantitative real time PCR）检测乳腺癌组织中PVT1、miR-214以及PD1基因的相对表达量；通过卡方检验分析不同PVT1表达水平的乳腺癌患者生存和预后之间的关联；通过慢病毒包装技术筛选PVT1过表达的乳腺癌细胞系；通过MTT实验检测PVT1过表达对乳腺癌细胞增殖的影响；通过荧光素酶报告基因实验检测miR-214对PVT1的结合；通过western blot实验检测PVT1过表达细胞系中PD1蛋白质的表达情况；通过MTT实验检测PVT1过表达细胞系对PD1抑制剂敏感性的影响；结果： PVT1在乳腺癌中表达上调（P<0.05），缩短乳腺癌患者生存期（P<0.01）；miR-214在乳腺癌患者中低表达（P=0.015），与PVT1的表达量成正相关（r=0.501，P<0.01）；过表达PVT1能够上调PD1基因（P<0.05）和蛋白质（P<0.05）的表达。通过荧光素酶报告基因实验发现miR-214能够靶向结合PVT1（P<0.05）；PVT1过表达促进乳腺癌细胞系增殖（P<0.05）降低乳腺癌细胞对PD1抑制剂敏感性。结论： miR-214靶向PVT1抑制乳腺癌细胞对PD1抑制剂敏感性。

Objective: To investigate the molecular mechanism of miR-214 targeting lnc-PVT1 to inhibit the sensitivity of breast cancer cells to PD1 inhibitors. Methods: The relative expression levels of PVT1,miR-214 and PD1 genes in breast cancer tissues were detected by quantitative real-time PCR. The chi-square test was used to analyze the correlation between survival and prognosis of breast cancer patients with different PVT1 expression levels. PVT1 overexpressed breast cancer cell lines were screened by lentivirus packaging. The effect of PVT1 overexpression on the proliferation of breast cancer cells was detected by MTT assay. Mir-214 binding to PVT1 was detected by luciferase reporter assay. The expression of PD1 protein in PVT1 overexpressed cell lines was detected by western blot. The sensitivity of PVT1 overexpressed cell lines to PD1 inhibitors was determined by MTT assay. Results: The expression of PVT1 was up-regulated in breast cancer (P<0.05),and the survival time of breast cancer patients was shortened (P<0.01). miR-214 was low in breast cancer patients (P=0.015) and positively correlated with the expression of PVT1 (R=0.501,P<0.01). Overexpression of PVT1 can up-regulate the expression of PD1 gene (P<0.05) and protein (P<0.05). The luciferase reporter gene assay showed that mir-214 could target PVT1 (P<0.05). Overexpression of PVT1 promoted the proliferation of breast cancer cell lines (P<0.05) and reduced the sensitivity of breast cancer cells to PD1 inhibitors. Conclusion: Mir-214 targets PVT1 to inhibit the sensitivity of breast cancer cells to PD1 inhibitors.

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