RNA干扰水牛MTPN基因对颗粒细胞增殖、凋亡及激素分泌的影响

doi:10.16431/j.cnki.1671-7236.2019.03.019

Abstract

Abstract:

This study was aimed to investigate the effect of MTPN gene knockdown on proliferation and apoptosis of buffalo granulosa cells in vitro and estrogen and progesterone secretion.The expression of MTPN gene in granulosa cells was knocked down by RNAi technology.The expression of MTPN gene and proliferation and cell cycle-related genes were detected by Real-time PCR,the cell proliferation and cell cycle distribution were detected by CCK-8 assay and flow cytometry,respectively.The estrogen and progesterone levels in the cell culture medium were determined by ELISA kit.The results showed that the relative expression of MTPN gene was decreased by 60% after siRNA (si-MTPN) transfection (P<0.01).The cell proliferation was significantly inhibited (P<0.05),the number of cells in G1 phase decreased,and the number of cells in S phase increased,the number of cells in G2 phase extremely significantly increased (P<0.01),and the cells were blocked in G2 phase.The expressions of Cyclin D2 and Cytochrom C genes were significantly up regulated (P<0.05),and the expressions of Caspase9 and Fas genes were extremely significantly up regulated (P<0.01).ELISA results showed that the levels of estrogen and progesterone secretion were significantly decreased (P<0.05).In conclusion,knockdown of MTPN gene could inhibit the proliferation of buffalo granulosa cells in vitro and the secretion of estrogen and progesterone by regulating the expression of related genes,which laid a reference for elucidating the molecular mechanism of MTPN gene involved in follicular development of livestock.

Key words: buffalo; MTPN gene; granulosa cells; proliferation; apoptosis; estrogn; progesterone

CLC Number:

S814.8

CHEN Chao, LI Jun, HUA Liping, AN Zhigao, HU Xiangwei, LIU Shenhe, YANG Liguo. Effect of RNA Interference Buffalo MTPN Gene on Proliferation, Apoptosis of Granulosa cells and Hormone Secretion[J]. , 2019, 46(3): 800-807.

References

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Effect of RNA Interference Buffalo MTPN Gene on Proliferation, Apoptosis of Granulosa cells and Hormone Secretion

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