雌激素受体对乳腺癌细胞截短型神经激肽受体-1的调控作用

目的探讨雌激素受体α（ERα）阳性的乳腺癌细胞中ERα对神经激肽受体-1截短型变异体（NK1R-Tr）的调控作用，以及ERα是否通过调控NK1R-Tr的表达，间接调控细胞的增殖能力。方法染色质免疫共沉淀（CHIP）实验验证ERα是否可以结合到NK1R-Tr启动子上游的ERα反应元件，直接调控NK1R-Tr的表达；荧光素酶报告基因实验验证ERα是否对NK1R-Tr的表达起正性调控作用。Westernblot实验和RT-PCR实验检测乳腺癌细胞系MCF-7和T47D的ERα和NK1R-Tr在蛋白水平和mRNA水平的表达情况；以及在ERα激动剂雌二醇（E2）刺激的条件下，小干扰RNA敲除ERα后，N...

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Bibliographic Details
Published in天津医药 Vol. 44; no. 12; pp. 1409 - 1413
Main Author 刘晓彬;仝颖娜;张露芳;周云丽
Format Journal Article
LanguageChinese
Published 天津医科大学肿瘤医院检验科，国家肿瘤临床医学研究中心，天津市恶性肿瘤临床医学研究中心，天津市“肿瘤防治”重点实验室 邮编300060 2016
Subjects
乳腺肿瘤;细胞系,肿瘤;雌激素受体α;受体,神经激肽1;RNA,小分子干扰;染色质免疫沉淀法;细胞增殖;荧光素酶报告基因
RNA,small interfering
receptors,neurokinin- 1
cell proliferation
luciferase reporter
breast neoplasms
荧光素酶报告基因
estrogen receptor alpha
cell line,tumor
chromatin immunoprecipitation
RNA,小分子干扰
乳腺肿瘤
染色质免疫沉淀法
雌激素受体α
细胞系,肿瘤
受体,神经激肽1
细胞增殖
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ISSN0253-9896
DOI10.11958/20161193

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Summary:目的探讨雌激素受体α（ERα）阳性的乳腺癌细胞中ERα对神经激肽受体-1截短型变异体（NK1R-Tr）的调控作用，以及ERα是否通过调控NK1R-Tr的表达，间接调控细胞的增殖能力。方法染色质免疫共沉淀（CHIP）实验验证ERα是否可以结合到NK1R-Tr启动子上游的ERα反应元件，直接调控NK1R-Tr的表达；荧光素酶报告基因实验验证ERα是否对NK1R-Tr的表达起正性调控作用。Westernblot实验和RT-PCR实验检测乳腺癌细胞系MCF-7和T47D的ERα和NK1R-Tr在蛋白水平和mRNA水平的表达情况；以及在ERα激动剂雌二醇（E2）刺激的条件下，小干扰RNA敲除ERα后，NK1R-Tr在不同水平的表达情况；小干扰RNA敲除NK1R-Tr后，CCK-8和克隆形成实验检测敲除NK1R-Tr的乳腺癌细胞的增殖能力。结果在NK1R-Tr基因启动子上游存在ERα的反应元件，ERα在E2存在条件下作用于该反应元件，对NK1R-Tr的表达起正性调控作用。同样在E2刺激的条件下，敲除乳腺癌细胞MCF-7内源性ERα后，NK1R-Tr在蛋白水平和mRNA水平的表达均下降；且敲除NK1R-Tr的MCF-7细胞增殖能力较未敲除组明显降低。结论在ERα阳性的乳腺癌细胞中，ERα正性调控NK1R-Tr的表达，从而增强细胞的增殖能力。
Bibliography:breast neoplasms; cell line, tumor; estrogen receptor alpha; receptors, neurokinin- 1; RNA, small interfering; chromatin immunoprecipitation; cell proliferation; luciferase reporter
To analyze the regulation of estrogen receptor α (ERα) on truncated neurokinin-1 receptor (NK1R-Tr), and the influence of this regulation on cell proliferation in estrogen receptor-positive breast cancer cell lines.Methods The chromatin immune coprecipitation (CHIP) was used to observe the transcriptional regulation function of ERαon NK1R- Tr in breast cancer cells. Luciferase reporter gene assay was used to verify whether ERα played a positiveregulatory role in the expression of NK1R-Tr. Western blot assay and real-time-PCR were used to detect the expression ofERα and NK1R-Tr in breast cancer cells, MCF-7 and T47D, as well as the expression of NK1R-Tr protein and mRNAlevel. NK1R-Tr levels were also detected after using estradiol (E2, ERα agonist) and small interfering RNA (knock outERα). CCK- 8 and clone formation experimen were u
ISSN:0253-9896
DOI:10.11958/20161193