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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| Published in | 天津医药 Vol. 44; no. 12; pp. 1409 - 1413 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| 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
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0253-9896 |
| DOI | 10.11958/20161193 |
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| Abstract | 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。 |
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| AbstractList | R737.9; 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。Western blot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。 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。 |
| Abstract_FL | Objective 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 positive regulatory role in the expression of NK1R-Tr. Western blot assay and real-time-PCR were used to detect the expression of ERα and NK1R-Tr in breast cancer cells, MCF-7 and T47D, as well as the expression of NK1R-Tr protein and mRNA level. NK1R-Tr levels were also detected after using estradiol (E2, ERα agonist) and small interfering RNA (knock out ERα). CCK-8 and clone formation experimen were used to detect the proliferation ability of breast cancer cells after knocking out NK1R-Tr with small interfering RNAs. Results CHIP test and Luciferase reporter gene assay proved that ERα can positively regulate the expression of NK1R-Tr via the ERα sequences in the upstream of the NK1R-Tr gene promoter. The expression of NK1R-Tr at both protein level and mRNA level dropped in the estrogen receptor-positive breast cancer cell line MCF-7 upon knocking out ERα. After knocking out NK1R-Tr, the proliferation ability of estrogen receptor-positive breast cancer cells was lower than that of the control group. Conclusion The ERα positively regulates the expression of NK1R-Tr, resulting in the increased cell proliferation in estrogen positive breast cancer cells. |
| Author | 刘晓彬;仝颖娜;张露芳;周云丽 |
| AuthorAffiliation | 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,300060 |
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| Author_FL | LIU Xiaobin ZHANG Lufang TONG Yingna ZHOU Yunli |
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| DocumentTitleAlternate | Study on the regulation of ERα on NK1R-Tr in breast cancer cells |
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| Keywords | 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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| Notes | 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 |
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| Publisher | 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 |
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| SubjectTerms | 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 |
| Title | 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 |
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