猪RPL36A基因对PK15细胞增殖过程的影响

doi:10.16431/j.cnki.1671-7236.2023.08.001

Abstract

Abstract: 【Objective】 The purpose of this study was to investigate the effect of ribosomal protein L36A (RPL36A) gene on PK15 cells proliferation,and lay the foundation for the analysis of the physiological mechanism of the difference in growth rate between Mashen and Large White pigs.【Method】 siRNA and overexpression vectors of RPL36A gene were constructed and transfected into PK15 cells by liposome transfection method.The expression efficiency of RPL36A gene and the expression of genes (PCNA,Ki67,Cyclin B and CDK4) related cell proliferation in PK15 cells were detected by Real-time quantitative PCR and Western blotting.The scratch tests,CCK-8 and EdU methods were used to detect cell proliferation.【Result】 After overexpression of RPL36A gene in PK15 cells,the expression of PCNA,Ki67 and CDK4 genes mRNA were extremely significant increased (P＜0.01),the expression of Cyclin B gene mRNA was significantly increased (P＜0.05),and the expression of PCNA protein was significantly increased (P＜0.05).The number of PK15 cells at 48 h was extremely significantly higher than control group (P＜0.01),and the rate of proliferation was increased.The number of positive cells of PK15 cells were extremely significantly increased (P＜0.01).After interference of RPL36A gene in PK15 cells,the expression of PCNA and Cyclin B genes mRNA were extremely significant decreased (P＜0.01),the expression of Ki67 and CDK4 genes mRNA were significantly decreased (P＜0.05),and the expression of PCNA protein was significantly decreased (P＜0.05).The number of PK15 cells at 48 h was significantly lower than siNC group (P＜0.05),and the rate of proliferation was decreased.The number of positive cells was extremely significantly decreased (P＜0.01).【Conclusion】 After overexpression and interference of RPL36A gene in PK15 cells,the expression of PCNA,Ki67,Cyclin B and CDK4 genes,which were key genes for cell proliferation,as well as the number of PK15 cells and the number of positive cells,were significantly altered.The proliferation of PK15 cells was affected by RPL36A gene.

Key words: pigs; RPL36A gene; PK15 cells; overexpression; interference; cell proliferation

CLC Number:

S828

WANG Shouyuan, YUN Hongmei, SHI Mingyue, QIN Yunmeng, LI Xiong, CHEN Junzhou, ZHOU Chenbo, CAO Guoqing. Effect of Porcine RPL36A Gene on PK15 Cell Proliferation[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(8): 3035-3044.

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Effect of Porcine RPL36A Gene on PK15 Cell Proliferation

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