Study on the regulation of ERαon NK1R-Tr in breast cancer cells
10.11958/20161193
- VernacularTitle: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
- Author:
Xiaobin LIU
;
Yingna TONG
;
Lufang ZHANG
;
Yunli ZHOU
- Keywords:
breast neoplasms;
cell line,tumor;
estrogen receptor alpha;
receptors,neurokinin- 1;
RNA,small interfering;
chromatin immunoprecipitation;
cell proliferation;
luciferase reporter
- From:
Tianjin Medical Journal
2016;44(12):1409-1413
- CountryChina
- Language:Chinese
-
Abstract:
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.
