<em>NONHSAT184603.1</em>基因对解旋酶基因<em>BLM</em>表达的影响

doi:10.3969/j.issn.1674-7968.2020.09.013

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摘要解旋酶BLM (bloom helicase)在细胞的增殖生长过程中有着重要的调控作用,YAP (yes associated protein)和TAZ (transcriptional coactivator with PDZ-binding motif)蛋白作为哺乳动物Hippo信号通路下游的转录共激活因子和关键成分,参与调控细胞的增殖、凋亡以及迁移等多种生理过程,而长链非编码RNA (long non-coding RNA, LncRNA)在细胞的生长增殖过程中具有重要的调控作用。为探究LncRNA对BLM、YAP、TAZ表达的影响,本研究通过mirBase (http://www.mirbase.org/)、NONCODE (http://www.noncode.org/)及自主开发程序SWChen 2.3筛选与BLM解旋酶相关的LncRNAs,利用荧光定量PCR技术检测筛选的LncRNA在22RV-1、PC3、LNCap、WPMY-1四种细胞系中的表达情况,构建LncRNA超表达载体,检测转染后24及48 h LncRNA对BLM、YAP及TAZ的影响。本研究以筛选出的NONHSAT184603.1 (缩写为N183)(GenBank No. m121218_085412_00126_c100)为候选基因,荧光定量PCR检测结果显示,N183在前列腺癌细胞系22RV-1和LNCap细胞中的表达极显著低于前列腺正常上皮细胞系WPMY-1 (P<0.01),但在前列腺癌细胞系PC3中的表达极显著高于WPMY-1 (P<0.01);WPMY-1细胞转染N183超表达载体24 h后与对照组比较,BLM表达极显著下调(P<0.01),且TAZ表达也是极显著下调(P<0.01),转染48 h后BLM和YAP表达均极显著上调(P<0.01);PC3细胞在转染48 h后,BLM表达量显著下调(P<0.05),但YAP和TAZ表达差异不显著(P>0.05);以上结果表明N183在前列腺癌细胞及正常细胞中能够抑制BLM基因的表达,但对增殖指标基因YAP和TAZ的影响具有差异,为今后以BLM解旋酶为抗癌靶标对抗癌症提供基础数据。

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	杨丽红
	吴小敏
	崔小芝
	吴巧群
	罗斌杰
	谌颖莲
	许厚强

关键词 ：前列腺癌细胞, BLM解旋酶, Hippo信号通路, NONHSAT184603.1

Abstract：BLM (bloom helicase) plays an important role in cell proliferation and growth. YAP (yes associated protein) and TAZ (transcriptional coactivator with PDZ-binding motif) porteins, as transcriptional coactivators and key components downstream of the mammalian Hippo signal pathway, are involved in the regulation of cell proliferation, apoptosis, migration and other physiological processes.And the long non-coding RNA (LncRNA) is very critical for regulating gene expression in cell growth and proliferation. To explore LncRNAs effect on gene expression of BLM, YAP, TAZ and screen BLM helicase related LncRNAs, fluorescence quantitative PCR technique had been used to detect the expression of selected LncRNA in 4 kinds of cell lines (22RV-1, PC3, LNCap, WPMY-1). Construction of LncRNA over-expression vector had been transfected in PC3 cells and WPMY-1 to detect the impact on BLM, YAP and TAZ gene after transfection LncRNA 24 and 48 h by using the fluorescent quantitative PCR technique. The NONHSAT184603.1 (abbreviated as N183) (GenBank No. m121218_085412_00126_c100) was successfully screened as candicate. The fluorescence quantitative PCR results showed that the expression of N183 in 22RV-1 and LNCap cells was significantly lower than that in normal prostate epithelial cell line WPMY-1 (P<0.01), and the expression in PC3 cells was significantly higher than that in WPMY-1 (P<0.01). After transfection with N183 over-expression vector, compared with the control group, the expression of BLM and TAZ were significantly down-regulated 24 h after transfection (P<0.01), and the expression of BLM and YAP were significantly up-regulated 48 h after transfection (P<0.01). BLM expression was significantly down-regulated (P<0.05), but there were no significant difference in YAP and TAZ expression in PC3 cells 48 h after transfection with N183 over-expression vector (P>0.05).The above results indicated that N183 could inhibit BLM gene expression in prostate cancer cells and normal cells but had different effect no YAP and TAZ genes, which made a contribution of basic data for anti-cancer researches by targeting BLM helicase.

Key words： Prostate cancer cell BLM helicase Hippo signal pathway NONHSAT184603.1

收稿日期: 2020-02-16

ZTFLH:	S828
	Q28

通讯作者: *,gzdxxhq@163.com

引用本文:

杨丽红, 吴小敏, 崔小芝, 吴巧群, 罗斌杰, 谌颖莲, 许厚强. NONHSAT184603.1基因对解旋酶基因BLM表达的影响[J]. 农业生物技术学报, 2020, 28(9): 1642-1651.
YANG Li-Hong, WU Xiao-Min, CUI Xiao-Zhi, WU Qiao-Qun, LUO Bin-Jie, CHEN Ying-Lian, XU Hou-Qiang. Effect of NONHSAT184603.1 Gene on the Expression of Helicase Gene BLM. 农业生物技术学报, 2020, 28(9): 1642-1651.

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http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2020.09.013 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2020/V28/I9/1642

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