miR-16对人乳腺癌细胞MDA-MB-231增殖和凋亡的作用及机制

doi:10.20047/j.issn1673-7210.2023.25.06

摘要
图/表
参考文献(24)
相关文章 (15)

全文: PDF (1035 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要目的探究miR-16对人乳腺癌细胞MDA-MB-231增殖和凋亡的作用及机制。方法体外培养细胞，分组如下：人乳腺上皮细胞HBL-100（HBL-100组）和人乳腺癌细胞MDA-MB-231（MDA-MB-231组）。采用荧光实时定量PCR检测miR-16表达，蛋白质印迹法检测β2肾上腺素受体（β2-AR）蛋白表达。细胞瞬时转染，分组如下：MDA-MB-231组，MDA-MB-231+miR-16拟似物（miR-16 mimics组），MDA-MB-231+miR-16抑制物（miR-16 inhibitor组），MDA-MB-231+阴性对照（阴性对照组）。MTT比色法和Annexin V-FITC双染流式细胞仪检测观察miR-16对MDA-MB-231细胞增殖和凋亡影响。双萤光素酶报告基因验证miR-16与β2-AR靶向关系。结果 MDA-MB-231组miR-16表达低于HBL-100组，β2-AR蛋白表达高于HBL-100组，差异有统计学意义（P＜0.05）。MDA-MB-231组与阴性对照组细胞增殖比较，差异无统计学意义（P＞0.05）。miR-16 mimics组细胞增殖低于MDA-MB-231组，miR-16 inhibitor组细胞增殖高于MDA-MB-231组，差异有统计学意义（P＜0.05）。MDA-MB-231组与阴性对照组细胞凋亡率比较，差异无统计学意义（P＞0.05）。miR-16 mimics组细胞凋亡率高于MDA-MB-231组，miR-16 inhibitor组细胞凋亡率低于MDA-MB-231组，差异有高度统计学意义（P＜0.01）。双萤光素酶报告基因结果显示，与MDA-MB-231组比较，miR-16 inhibitor组和阴性对照组萤光素酶活性都没有明显变化，差异无统计学意义（P＞0.05）。与MDA-MB-231组比较，具有β2-AR的3’UTR序列的miR-16 mimics组细胞的萤光素酶活性降低，差异有高度统计学意义（P＜0.01）；与MDA-MB-231组比较，具有突变的β2-AR的3’UTR序列的miR-16 mimics组细胞萤光素酶活性无明显改变，差异无统计学意义（P＞0.05）。结论 miR-16可能通过靶向β2-AR调控乳腺癌细胞增殖与凋亡。

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	刘嘉祺　　周福波　　林峰　　付惠

关键词 ： miR-16, &beta, 2肾上腺素受体, MDA-MB-231, 增殖, 凋亡

Abstract：Objective To investigate the effect and mechanism of miR-16 on proliferation and apoptosis of human breast cancer cells MDA-MB-231. Methods The cells cultured in vitro were divided into the following groups: human mammary epithelial cells HBL-100 (HBL-100 group ) and human breast cancer cells MDA-MB-231 (MDA-MB-231 group ). Real-time fluorescent quantitative PCR was used to detect miR-16 expression, and Western blot was used to detect β2 adrenergic receptor (β2-AR) protein expression. Transient transfection of cells was grouped as follows: MDA-MB-231 group, MDA-MB-231+miR-16 mimics (miR-16 mimics group), MDA-MB-231+miR-16 inhibitor (miR-16 inhibitor group), MDA-MB-231+ negative control (negative control group). The effects of miR-16 on proliferation and apoptosis of MDA-MB-231 cells were observed by MTT colorimetry and Annexin V-FITC double dye flow cytometry. The targeting relationship between miR-16 and β2-AR was verified by dual luciferase reporter genes. Results The expression of miR-16 in MDA-MB-231 group was lower than that in HBL-100 group, and the expression of β2-AR protein was higher than that in HBL-100 group, and the differences were statistically significant (P＜0.05). There was no significant difference in cell proliferation between MDA-MB-231 group and negative control group (P＞0.05). Cell proliferation in miR-16 mimics group was lower than that in MDA-MB-231 group, and cell proliferation in miR-16 inhibitor group was higher than that in MDA-MB-231 group, and the differences were statistically significant (P＜0.05). There was no significant difference in apoptosis rate between MDA-MB-231 group and negative control group (P＞0.05). The apoptosis rate in miR-16 mimics group was higher than that in MDA-MB-231 group, and the apoptosis rate in miR-16 inhibitor group was lower than that in MDA-MB-231 group, and the differences were highly statistically significant (P＜0.01). The results of dual luciferase reporter gene showed that, compared with MDA-MB-231 group, luciferase activity in miR-16 inhibitor group and negative control group had no significant changes, with no statistical significance (P＞0.05). Compared with MDA-MB-231 group, the luciferase activity of miR-16 mimics group with β2-AR 3’UTR sequence decreased, the difference was highly statistically significant (P＜0.01). Compared with MDA-MB-231 group, there was no significant change in luciferase activity in miR-16 mimics group with mutated β2-AR 3’UTR sequence, with no statistical significance (P＞0.05). Conclusion miR-16 may regulate the proliferation and apoptosis of breast cancer cells by targeting β2-AR.

Key words： miR-16 β2-adrenergic receptor MDA-MB-231 Proliferation Apoptosis

基金资助:黑龙江省省属高校基本科研业务费项目（2018-KYYWF-0144）

作者简介: 刘嘉祺（1981-），女，博士，主要从事药理学教学与科研工作。

引用本文:

刘嘉祺　　周福波　　林峰　　付惠. miR-16对人乳腺癌细胞MDA-MB-231增殖和凋亡的作用及机制[J]. 中国医药导报, 2023, 20(25): 38-42.
LIU Jiaqi ZHOU Fubo LIN Feng FU Hui. Effect and mechanism of miR-16 on proliferation and apoptosis of human breast cancer cells MDA-MB-231. 中国医药导报, 2023, 20(25): 38-42.

链接本文:

https://www.yiyaodaobao.com.cn/CN/10.20047/j.issn1673-7210.2023.25.06 或 https://www.yiyaodaobao.com.cn/CN/Y2023/V20/I25/38

[1] Liang Y，Song X，Li Y，et al. A novel long non-coding RNA- PRLB acts as a tumor promoter through regulating miR- 4766-5p/SIRT1 axis in breast cancer [J]. Cell Death Dis，2018， 9（5）：563.
[2] 宋莉，戴小斌，贾琦，等. β-肾上腺素能受体在乳腺癌发生发展中的作用[J].中国药理学通报，2021，37（3）：309 -313.
[3] Cindy K，Robert L，Erica K，et al. The β2-adrenoceptor activates a positive cAMP-calcium feedforward loop to drive breast cancer cell invasion [J]. FASEB J，2016，30（3）：1144-1154.
[4] Xiao ML，Yi Z，Yu EN，et al. miR-16-5p Inhibits Malignant Progression of Cervical Cancer by Targeting YAP1 [J]. Onco Targets Ther，2020，30（13）：931-944.
[5] Yong L，Dong LT，Shi YZ，et al. Serum microRNA-195 as a potential diagnostic biomarker for breast cancer： a systematic review and meta-analysis [J]. Int J Clin Exp Pathol，2019， 12（11）：3982-3991.
[6] 徐凯，金延玲，刘伟，等.抗氧化剂N-乙酰半胱氨酸对人乳腺癌细胞MDA-MB-231迁移能力的影响[J].兰州大学学报（医学版），2021，47（4）：30-34.
[7] Liu J Q，Sun F，Wang YY，et al. Suppression of microRNA- 16 protects against acute myocardial infarction by reversing beta2-adrenergic receptor down-regulation in rats [J]. Oncotarget，2017，8（12）：20122-20132.
[8] 骆广涛，王本忠.miRNA-4465过表达对乳腺癌 MDA-MB- 231细胞迁移侵袭的影响及其机制[J].安徽医科大学学报，2015，50（11）：1570-1574.
[9] Baharuddin AA，Roosli RAJ，Zakaria ZA，et al. Dicranopteris linearis extract inhibits the proliferation of human breast cancer cell line（MDA-MB-231） via induction of S-phase arrest and apoptosisr [J]. Pharm Biol，2018，56（1）：422-432.
[10] 钱洁，谭佳妮，程海波，等.小檗碱对乳腺癌MDA-MB- 231细胞增殖抑制作用及其分子机制研究[J].天然产物研究与开发，2019，31（3）：422-427.
[11] Wang ZM，Hu SY，Li XY，et al. MiR-16-5p suppresses breast cancer proliferation by targeting ANLN [J]. BMC Cancer，2021，21：1188.
[12] Duffy MJ，Maguire TM，Hill A，et al. Metalloproteinases：role in breast carcinogenesis，invasion and metastasis [J]. Breast Cancer Res，2000，2（4）：252-257.
[13] Steeg PS. Molecular biology of breast cancer metastasis. ‘Has it spread?’：disarming one of the most terrifying questions [J]. Breast Cancer Res，2000，2（6）：396-399.
[14] Bartel DP. MicroRNAs：target recognition and regulatory functions [J]. Cell，2009，136：215-233.
[15] Fabian MR，Sonenberg N，Filipowicz W. Regulation of mRNA translation and stability by microRNAs [J]. Annu Rev Biochem，2010，7（9）：351-379.
[16] Lerner M，Harada M，Loven J，et al. DLEU2，frequently deleted in malignancy，functions as a critical host gene of the cell cycle inhibitory microRNAs miR-15a and miR- 16-1 [J]. Exp Cell Res，2009，315：2941-2952.
[17] Ambreen U，Amir A，Bushra S，et al. MiR-16：A novel hereditary marker in breast cancer and their offspring [J]. J Pak Med Assoc，2017，67（3）：446-450.
[18] Tang X，Jin L，Cao P，et al. MicroRNA-16 sensitizes breast cancer cells to paclitaxel through suppression of IKBKB expression [J]. Oncotarget，2016，7（17）：23668-23683.
[19] Guo X，Connick MC，Vanderhoof J，et al. MicroRNA-16 modulates HuR regulation of cyclin E1 in breast cancer cells [J]. Int J Mol Sci，2015，16（4）：7112-7132.
[20] Chen D，Xing W，Hong J，et al. The beta2-adrenergic receptor is a potential prognostic biomarker for human hepatocellular carcinoma after curative resection [J]. Ann Surg Oncol，2012，19（11）：3556-3565.
[21] Monique B，Xiu NL，John VH. β-Adrenergic Signaling in Lung Cancer：A Potential Role for Beta-Blockers [J]. J Neuroimmune Pharmacol，2020，15（1）：27-36.
[22] Zhen L，Wei YY，Ying Z. Beta-blocker and survival in patients with lung cancer：Ameta-analysis [J]. PLoS One，2021， 16（2）：e0245773.
[23] Choy C，Raytis JL，Smith DD，et al. Inhibition of β2-adrenergic receptor reduces triple-negative breast cancer brain metastases：The potential benefit of perioperative β-blockade [J]. Oncol Rep，2016，35（6）：3135-3142.
[24] Qin JF，Jin FJ，Li N，et al. Adrenergic receptor β2 activation by stress promotes breast cancer progression through mac- rophages M2 polarization in tumor microenvironment [J]. BMB Rep，2015，48（5）：295-300.