胸腺上皮细胞中雌激素诱导lncRNA的鉴定分析及表达载体构建

doi:10.16431/j.cnki.1671-7236.2021.01.007

Abstract

Abstract: To study the regulation role of estradiol (E₂) on long non-coding RNA (lncRNA) expression in thymic epithelial cells (TEC).The cell phenotypic changes were observed after the 50 nmol/L E₂ was added in cultured mouse epithelial cell line 1 (MTEC1),while the cell viability was also measured with CCK-8 kit.Furthermore,total RNA was extracted and the expression regulatory role of E₂ on lncRNA-2410006H16Rik was verified by Real-time quantitative PCR.Moreover,the target gene was amplified by RT-PCR to construct a recombinant vector of pEGFP-N1-lncRNA-2410006H16Rik.The results showed the proliferation of MTEC1 cells was significantly inhibited by 50 nmol/L E₂,and the D_{450 nm} value was also extremely significantly reduced as compared with control group detected by CCK-8 (P<0.01),indicating that the cell viability was extremely significantly decreased by E₂.Real-time quantitative PCR results indicated that the expression of lncRNA-2410006H16Rik in MTEC1 cells was extremely significantly up-regulated by 50 nmol/L E₂ (P<0.01),and about two fold-changes higher than control group cells,which was consistent with the high-throughput sequencing results.RT-PCR,double enzyme digestion and sequencing results showed that the pEGFP-N1-lncRNA-2410006H16Rik expression vector was successfully constructed.All the above results indicated that the expression of lncRNA-2410006H16Rik in TECs was closely related to the effect of E₂,which laid a foundation for further verification of the regulatory function of lncRNA-2410006H16Rik at the cellular level.

Key words: lncRNA; lncRNA-2410006H16Rik; E₂; MTEC1; expression vector

CLC Number:

Q291

GUO Dongguang, CHEN Mingyan, CUI Fangwei, LI Wenming, GUO Zanying, ZHU Yanping, YUE Feng, WANG Xuannian. Identification Analysis and Construction of Expression Vector of Estrogen-induced lncRNA in Thymic Epithelial Cells[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 64-71.

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Identification Analysis and Construction of Expression Vector of Estrogen-induced lncRNA in Thymic Epithelial Cells

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