雌激素通过介导HOTAIR表达对子宫内膜癌细胞增生及裸鼠致瘤能力的影响

doi:10.3969/j.issn.1006-7795.2019.06.016

摘要/Abstract

摘要： 目的探讨雌激素是否通过介导HOTAIR表达对子宫内膜癌细胞增生及裸鼠致瘤能力产生影响。方法构建慢病毒介导干扰HOTAIR表达的shHOTAIR及阴性对照shNC稳定转染的子宫内膜癌Ishikawa细胞株，qRT-PCR检测HOTAIR的敲减水平。实验细胞分为4组：对照组（Ishikawa细胞）、雌二醇（17β-estradiol，E2）组（E2+Ishikawa细胞）、E2+shNC组（E2+shNC细胞）、E2+shHOTAIR组（E2+shHOTAIR细胞），qRT-PCR检测E2对4组细胞中HOTAIR表达的影响。构建E2作用于HOTAIR表达不同的4组Ishikawa细胞的裸鼠移植瘤模型，记录成瘤时间、成瘤率及移植瘤生长曲线，剥离移植瘤，测量体积及质量，组织切片病理学检测。结果成功构建慢病毒介导干扰HOTAIR表达的shHOTAIR及阴性对照shNC稳定转染的Ishikawa细胞株，HOTAIR表达被有效敲减（P<0.001）；E2组细胞中HOTAIR的相对表达量明显高于对照组（P<0.001），E2+shHOTAIR组明显低于E2+shNC组（P<0.001）；成功构建E2作用于HOTAIR表达不同的4组Ishikawa细胞的裸鼠移植瘤模型；裸鼠致瘤能力对比：E2组>对照组，E2+shNC组>E2+shHOTAIR组；E2组移植瘤的终体积（P<0.001）及终质量（P<0.001）明显高于对照组，E2+shHOTAIR组移植瘤的终体积（P<0.01）及终质量（P<0.001）明显低于E2+shNC组；移植瘤HE染色符合子宫内膜癌细胞病理学特征。结论雌激素通过上调HOTAIR表达促进子宫内膜癌细胞的增生及裸鼠致瘤能力，有望为子宫内膜癌的治疗提供新靶点。

关键词: 子宫内膜癌, 雌激素, HOTAIR, 长链非编码RNA

Abstract: Objective To investigate the effect of estrogen on the proliferation and nude mouse tumorigenicity of endometrial carcinoma cells by mediating the expression of HOTAIR. Methods Lentivirus-mediated interference of HOTAIR expression shHOTAIR and negative control shNC were constructed and transfected into Ishikawa cells, the expression of HOTAIR was detected by qRT-PCR. Four groups were set up:control group (Ishikawa cells), E2 group (E2+Ishikawa cells), E2+shNC group (E2+shNC cells) and E2+shHOTAIR group (E2+shHOTAIR cells). qRT-PCR was used to detect the effect of E2 on the expression of HOTAIR in the four groups. The xenograft tumor model of nude mice treated with E2 and four groups Ishikawa cells of different expression of HOTAIR was constructed. The tumor formation time, the tumor formation rate and the growth curve of the xenograft tumor were recorded. The xenograft tumor was removed, whose volume and weight were measured. The pathological examination of the xenograft tumor was performed. Results The stable transfected cell lines with lentivirus-mediated interference of HOTAIR expression shHOTAIR and negative control shNC were successfully constructed, and the expression of HOTAIR was effectively inhibited (P<0.001). The relative expression of HOTAIR in E2 group was significantly higher than that of control group (P<0.001), and E2+shHOTAIR group was significantly lower than E2+shNC group (P<0.001). The xenograft tumor model of nude mice treated with E2 and four groups Ishikawa cells of different expression of HOTAIR was successfully constructed. The comparison of tumorigenic ability was as follows:E2 group>control group,E2+shNC group>E2+shHOTAIR group. The final volume (P<0.001) and final weight (P<0.001) of the xenograft tumor in E2 group were significantly higher than those in control group, and the final volume (P<0.01) and final weight (P<0.001) in E2+shHOTAIR group were significantly lower than those in E2+shNC group. The HE staining of the xenograft tumor was consistent with the pathological characteristics of endometrial carcinoma cells. Conclusion Estrogen promotes the proliferation and nude mouse tumorigenicity of endometrial carcinoma cells by up-regulating the expression of HOTAIR, which is expected to provide a new target for the treatment of endometrial carcinoma.

Key words: endometrial carcinoma, estrogen, HOTAIR, long non-coding RNAs

中图分类号:

R737.33

马旭兰, 夏迪, 王慧霄, 蒋子雯, 代荫梅. 雌激素通过介导HOTAIR表达对子宫内膜癌细胞增生及裸鼠致瘤能力的影响[J]. 首都医科大学学报, 2019, 40(6): 894-900.

Ma Xulan, Xia Di, Wang Huixiao, Jiang Ziwen, Dai Yinmei. Effect of estrogen on the proliferation and nude mouse tumorigenicity of endometrial carcinoma cells by mediating the expression of HOTAIR[J]. Journal of Capital Medical University, 2019, 40(6): 894-900.

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