B7-H3基因敲降ES-2细胞系构建及其对细胞增生的影响

doi:10.3969/j.issn.1006-7795.2021.01.011

首都医科大学学报 ›› 2021, Vol. 42 ›› Issue (1): 66-70.doi: 10.3969/j.issn.1006-7795.2021.01.011

B7-H3基因敲降ES-2细胞系构建及其对细胞增生的影响

邓梦琪, 张艳芹, 常翔宇, 吴迪, 徐春玉, 苗劲蔚^*

首都医科大学附属北京妇产医院妇瘤科,北京 100006

收稿日期:2020-10-23 出版日期:2021-02-21 发布日期:2021-02-02
基金资助:
北京市自然科学基金(7162063)。

Construction of B7-H3 knockdown ES-2 cell line and its effect on cell proliferation

Deng Mengqi, Zhang Yanqin, Chang Xiangyu, Wu Di, Xu Chunyu, Miao Jinwei^*

Department of Gynecologic Oncology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100006,China

Received:2020-10-23 Online:2021-02-21 Published:2021-02-02
Contact: ^*E-mail:jinweimiao@ccmu.edu.cn
Supported by:
Natural Science Foundation of Beijing(7162063)

摘要/Abstract

摘要： 目的构建B7同系物3(B7 homolog 3,B7-H3)敲降的ES-2细胞系并探究B7-H3基因对ES-2细胞系增生的影响。方法利用慢病毒介导的短发夹RNA(short hairpin RNA,shRNA)干扰技术在ES-2细胞中敲降B7-H3,利用荧光显微观察、实时荧光定量PCR 技术(real-time fluorescent quantitative PCR,qRT-PCR)及流式细胞术验证B7-H3敲降效果,并利用细胞计数试剂盒(cell counting kit-8,CCK-8)检验细胞活性改变。结果感染不同的shRNA,可呈现不同程度的B7-H3降低,CCK-8检测结果显示,敲除B7-H3后细胞增生速率明显低于对照组,差异有统计学意义(P<0.05)。结论使用慢病毒介导的shRNA干扰技术能成功构建B7-H3敲降的ES-2细胞系,B7-H3敲降后ES-2细胞增生速率显著降低。

关键词: ES-2细胞系, B7-H3, shRNA干扰技术, 细胞增生

Abstract: Objective To construct a B7-H3 knockdown ES-2 cell line and determine the effect of B7-H3 gene on the proliferation of ES-2 cell line. Methods Lentivirus mediated shRNA interference was used to establish ES-2 cell line with B7-H3 knockdown. Fluorescence microscopy,real-time fluorescent quantitative PCR(qRT-PCR) and flow cytometry were used to verify the effect of B7-H3 knockdown, and cell counting kit-8 (CCK-8) was used to test its effect on proliferation. Results The results of CCK-8 showed that the proliferation rate of the experimental group was significantly lower than that of the control group, with statistical significance(P<0.05). Conclusion B7-H3 knockdown ES-2 cell line is successfully constructed, and it is found that B7-H3 knockdown can significantly reduce the proliferation rate of ES-2 cells.

Key words: ES-2 cell line, B7-H3, shRNA interference technology, cell proliferation

中图分类号:

R713.6

邓梦琪, 张艳芹, 常翔宇, 吴迪, 徐春玉, 苗劲蔚. B7-H3基因敲降ES-2细胞系构建及其对细胞增生的影响[J]. 首都医科大学学报, 2021, 42(1): 66-70.

Deng Mengqi, Zhang Yanqin, Chang Xiangyu, Wu Di, Xu Chunyu, Miao Jinwei. Construction of B7-H3 knockdown ES-2 cell line and its effect on cell proliferation[J]. Journal of Capital Medical University, 2021, 42(1): 66-70.

参考文献

[1] Walker J, Powell B, Chen L, et al.Society of Gynecologic Oncology recom mendations for the prevention of ovarian cancer[J].Cancer, 2015, 7(121):2108-2120.
[2] Webb P, Green A, Jordan S, et al.Trends in hormone use and ovarian cancer incidence in US white and Australian women: implications for the future[J].Cancer Causes Control, 2017, 28(3):365-370.
[3] Cai D, Li J, Liu D, et al.Tumor-expressed B7-H3 mediates the inhibition of antitumor T-cell functions in ovarian cancer insensitive to PD-1 blockade therapy[J].Cell Mol Immunol, 2020, 17(3):227-236.
[4] Chen Y W, Tekle C, Fodstad O, et al.The immunoregulatory protein human B7H3 is a tumor- associated antigen that regulates tumor cell migration and invasion[J].Curr Cancer Drug Targets, 2008, 8(21):404-413.
[5] Yuan H, Wei X, Zhang G, et al. B7-H3 over expression in prostate cancer promotes tumor cell progression [J]. J Urolo, 2011, 186(3): 1093-1099.
[6] Lim S, Liu H, Arora R, et al. Immunoregulatory protein B7-H3 reprograms glucose metabolism in cancer cells by ROS-mediated stabilization of HIF-1α [J]. Cancer Res, 2016, 76(8): 2231-2242.
[7] Chien J, Poole E. Ovarian cancer prevention, screening and early detection: report from the 11th biennial ovarian cancer research symposium[J].Int J Gynecol Cancer, 2018, 27(49):S20-S22.
[8] 李宁, 孔北华, 尹如铁, 等. 晚期上皮性卵巢癌一线维持治疗专家共识[J].现代妇产科进展, 2019, 28(10):721-723.
[9] Disis M L, Taylor M H, Kelly K, et al. Efficacy and safety of avelumab for patients with recurrent or refractory ovarian cancer: phase 1b results from the JAVELIN solid tumor trial[J]. JAMA Oncol, 2019, 19(11):1468-1479.
[10]Pujade-Lauraine E, Fujiwara K, Dychter S S, et al. Avelumab (anti-PD-L1) in platinum-resistant/refractory ovarian cancer: JAVELIN ovarian 200 phase III study design[J]. Cancer Chemother Pharmacol, 2018,81(1): 17-38.
[11]Xu Y, Xiao Y, Luo C, et al. Blocking PD-1/PD-L1 by an ADCC enhanced anti-B7-H3/PD-1 fusion protein engages immune activation and cytotoxicity[J]. Int Immunopharmacol, 2020, 84:106584.
[12]Chen G, Tu Y, Aladelusi T O, et al. Knocking down B7H3 expression enhances cell proliferation of SHEDs via the SHP1/AKT signal axis[J]. Biochem Biophys Res Commun, 2020, 531(3):282-289.
[13]Liu F, Zhang T, Zou S, et al.B7-H3 promotes cell migration and invasion through the Jak2/Stat3/MMP9 signaling pathway in colorectal cancer[J].Mol Med Rep, 2015, 12(36):5455-5460.
[14]Liu H, Tekle C, Chen Y W, et al.B7-H3 silencing increases paclitaxel sensitivity by abrogating Jak2/Stat3 phosphorylation[J].Mol Cancer Ther, 2011, 10(69):960-971.
[15]Zhang T, Jiang B, Zou S T, et al.Overexpression of B7-H3 augments anti-apoptosis of colorectal cancer cells by Jak2-STAT3[J].World J Gastroenterol, 2015, 21(9):1804-1813.
[16]Tekle C, Nygren M K, Chen Y W, et al.B7-H3 contributes to the metastatic capacity of melanoma cells by modulation of known metastasis-associated genes[J].Int J Cancer, 2012, 130(9):2282-2290.
[17]Kang F B, Wang L, Jia H C, et al.B7-H3 promotes aggression and invasion of hepatocellular carcinoma by targeting epithelial-to-mesenchymal transition via JAK2/STAT3/Slug signaling pathway[J].Cancer Cell Int, 2015, 15(7):45.

B7-H3基因敲降ES-2细胞系构建及其对细胞增生的影响

Construction of B7-H3 knockdown ES-2 cell line and its effect on cell proliferation

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