干扰 LncRNA BCYRN1 对乳腺癌 MCF-7 细胞增殖、 运动和移植
瘤生长的影响

doi:10.3870/j.issn.1672-8009.2023.05.011

摘要/Abstract

摘要： 目的探讨短发卡 RNA (short hairpin RNA, shRNA) 干扰长链非编码 RNA BCYRN1 ( long non-coding RNA BCYRN1, LncRNA BCYRN1) 对乳腺癌 MCF-7 细胞增殖、运动及移植瘤生长的影响。方法实时定量 PCR (real-time quantitative PCR, RT-PCR) 检测乳腺癌及癌旁组织 BCYRN1 表达, 将乳腺癌 MCF-7 细胞分为空白对照组 (Control)、阴性对照组 ( shRNA-NC) 和干扰组 ( sh-BCYRN1), 通过试剂盒转染转入目的片段并进行 RT-PCR 验证, Edu 法、 transwell 法、划痕实验和蛋白质印迹法 (Western blotting) 分别检测细胞增殖、侵袭、迁移能力及其相关蛋白表达, 免疫荧光检测细胞波形蛋白 (Vimentin) 水平。通过裸鼠腋下注射 MCF-7 细胞建立裸鼠移植瘤模型, 待成瘤后将裸鼠分为空白对照组、干扰组, 每组各 20 只, 干扰组、空白对照组裸鼠分别给予 sh-BCYRN1 及其阴性对照转染干预, 观察 4 周后移植瘤 BCYRN1 表达、肿瘤重量和 Ki67、 N-钙粘蛋白 (N-cadherin) 蛋白表达。结果乳腺癌组织 BCYRN1 表达高于癌旁组织 (P< 0. 05)。比较空白对照组, 干扰组 Edu 阳性细胞百分比、单个视野细胞侵袭数目、细胞迁移率、 Vimentin 阳性率降低 (P< 0. 05), Ki67、 N-cadherin 蛋白表达降低, E-钙粘蛋白 (E-cadherin) 蛋白表达升高(P< 0. 05)。干扰组移植瘤 BCYRN1 表达降低, 肿瘤质量、 Ki67 及 N-cadherin 蛋白表达低于空白对照组移植瘤 (P< 0. 05)。结论 shRNA 干扰 LncRNA BCYRN1 可抑制乳腺癌 MCF-7 细胞增殖、运动和移植瘤生长。

关键词: 长链非编码 RNA, BCYRN1, MCF-7 乳腺癌细胞, 细胞增殖, 移植瘤

Abstract: Objective To explore the effect of LncRNA BCYRN1 on the proliferation and motility of MCF-7 breast cancer cells and the growth of xenograft in mice by using short hairpin RNA (shRNA) interfering. Methods The expression level of BCYRN1 in breast cancer and para-cancerous tissues was detected by real-time quantitative PCR (RT-PCR). MCF-7 cells were divided into 3 groups, the blank control group (Control), the negative control group (shRNA-NC) and the interference group (sh-BCYRN1). The target fragments were transfected, their effect on expressions of the target genes were then verified by RT-PCR. The cell proliferation, invasion, migration ability and the expression levels of related proteins were detected by Edu, transwell assay, wound-healing assay and Western blotting, respectively. Vimentin level was detected by immunofluorescence method. The nude mice were given subcutaneous injection of MCF-7 cells to construct xenograft models. After tumor formation, nude mice were divided into two groups, the blank control group (negative control ) and the interference group ( sh-BCYRN1 ), and there were 20 cases in each group. The tumor weight and the expression levels of BCYRN1, Ki67 and N-cadherin in the transplanted tumors were detected 4 weeks later. Results The expression level of BCYRN1 was higher in breast cancer tissues than in para-cancerous tissues ( P < 0. 05). The percentage of Edu positive cells, the number of migrated and invasion cells, the positive rate of Vimentin, and the expression levels of Ki67 and N-cadherin were decreased in the interference group when compared with the blank control group ( P < 0. 05), while the expression level of E-cadherin was increased ( P < 0. 05). In the interference group, the expression level of BCYRN1 was decreased, and the tumor mass, the expression levels of Ki67 and N-cadherin were lower than those in the blank control group (P< 0. 05). Conclusion Interfering of LncRNA BCYRN1 can inhibit the proliferation and motility of MCF-7 breast cancer cells and the growth of xenograft.

Key words: long non-coding RNA, BCYRN1, MCF-7 breast cancer cell, cell proliferation, xenograft

中图分类号:

R737. 9

吉棚 , 赵华山 , 王晨 , 刘淑荣 , 马建增. 干扰 LncRNA BCYRN1 对乳腺癌 MCF-7 细胞增殖、运动和移植瘤生长的影响[J]. 医学分子生物学杂志, 2023, 20(5): 444-449.

JI Peng , ZHAO Huashan , WANG Chen , LIU Shurong , MA Jianzeng . Effect of LncRNA BCYRN1 on Proliferation and Mobility of MCF-7 Breast Cancer Cells and Xenograft Growth in Mice[J]. Journal of Medical Molecular Biology, 2023, 20(5): 444-449.

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干扰 LncRNA BCYRN1 对乳腺癌 MCF-7 细胞增殖、运动和移植瘤生长的影响

Effect of LncRNA BCYRN1 on Proliferation and Mobility of MCF-7 Breast Cancer Cells and Xenograft Growth in Mice

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干扰 LncRNA BCYRN1 对乳腺癌 MCF-7 细胞增殖、 运动和移植 瘤生长的影响

Effect of LncRNA BCYRN1 on Proliferation and Mobility of MCF-7 Breast Cancer Cells and Xenograft Growth in Mice

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干扰 LncRNA BCYRN1 对乳腺癌 MCF-7 细胞增殖、运动和移植瘤生长的影响