黏蛋白16基因通过磷酸肌醇3-激酶/蛋白激酶B通路调节胆囊癌细胞活力和迁移

doi:10.16098/j.issn.0529-1356.2019.05.012

摘要/Abstract

摘要：

目的探讨黏蛋白16基因（MUC16）是否通过磷酸肌醇3-激酶/蛋白激酶B（PI3K/Akt）通路调节胆囊癌细胞（GBC-SD）活力和迁移。方法过表达MUC16后，通过Real-time PCR筛选相关基质金属蛋白酶（MMP）家族的蛋白质。其次，过表达MUC16、敲低MUC16和PI3K/Akt通路抑制剂BKM120处理后，通过免疫印迹法检测PI3K/Akt通路和MMP-9的蛋白水平。最后，过表达MUC16或敲低MUC16，通过细胞计数试剂盒8（CCK-8）和刮伤实验检测GBC-SD的活力和迁移情况。结果过表达MUC16后，MMP-9 mRNA的相对表达量显著升高（P<0.05）。过表达MUC16后， MMP-9、p-Akt、PI3K的蛋白水平明显升高（P<0.05），但是PI3K抑制剂BKM120可以避免这现象。而敲低MUC16后，发现MMP-9、p-Akt、PI3K的蛋白水平明显降低（P<0.05）。过表达MUC16后，GBC-SD的细胞活力和迁移能力明显增高（P<0.05），而敲低了MUC16后，GBC-SD的细胞活力和迁移能力明显降低（P<0.05）。另外，敲低MMP-9后，GBC-SD的细胞活力和迁移能力也明显降低（P<0.05）。但是，在过表达MUC16的同时敲低MMP-9，发现GBC-SD的细胞活力和迁移能力与对照组相比差异无显著性（P>0.05）。结论 MUC16激活PI3K/Akt通路促进MMP-9的蛋白表达，进而提升胆囊癌细胞GBC-SD的细胞活力和迁移能力。

关键词: 黏蛋白16基因, 磷酸肌醇3-激酶/蛋白激酶B通路, 基质金属蛋白酶9, 胆囊癌细胞, 免疫印迹法, 实时定量聚合酶链反应, 人

Abstract:

Objective To investigate the effect of mucin 16(MUC16) on the viability and migration of gallbladder cancer cells (GBC-SD) through phospho inositide 3-kinase/protein kinase B (PI3k/Akt) pathway. Methods Firstly, after overexpression of MUC16, the matrix metalloproteinase(MMP) family proteins were screened by Real-time PCR. Secondly, after overexpression of MUC16, knockdown of MUC16 and PI3K/Akt pathway inhibitor BKM120, the protein levels of PI3K/Akt pathway and MMP-9 were detected by Western blotting. Finally, the proliferation, viability and migration of gallbladder cancer cells were detected by cell counting kit-8 (CCK-8) and scratch test. Results After overexpression of MUC16, the relative expression of mRNA in MMP-9 increased significantly (P<0.05). After overexpression of MUC16, the levels of MMP-9, p-Akt and PI3K increased significantly (P<0.05), but PI3K inhibitor BKM120 could avoid this phenomenon. After knocking down MUC16, the protein levels of MMP-9, p-Akt and PI3K decreased significantly (P<0.05).After overexpression of MUC16, the cell viability and migration ability of GBC-SD increased significantly (P<0.05), while after knocking down MUC16, the cell viability and migration ability of GBC-SD decreased significantly (P<0.05). In addition, after knocking down MMP-9, the cell viability and migration ability of GBC-SD also decreased significantly (P<0.05). However, when MUC16 was overexpressed and MMP-9 was knocked down, there was no significant difference in cell viability and migration ability between GBC-SD and the control group (P>0.05). Conclusion MUC16 activates PI3K/Akt pathway to promote the expression of MMP-9 protein, thereby enhancing the cell viability and migration of GBC-SD.

Key words: Mucin 16, Phoshoinositide 3-kinase/protein kinase B pathway, Matrix metalloproteinase-9, Gallbladder cancer cell, Western blotting, Real-time PCR, Human

李新田蜜彭冰何莉莉 . 黏蛋白16基因通过磷酸肌醇3-激酶/蛋白激酶B通路调节胆囊癌细胞活力和迁移[J]. 解剖学报, 2019, 50(5): 613-619.

LI Xin TIAN Mi PENG Bing HE Li-li. Mucin 16 regulating activity and migration of gallbladder cancer cells through phosphoinositide 3-kinase/protein kinase B pathway[J]. Acta Anatomica Sinica, 2019, 50(5): 613-619.

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