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作者	单位	E-mail
金聪慧	南通大学附属肿瘤医院肿瘤内科江苏南通 226001	jch18862738535@163.com
苗波波	南通大学附属医院病理科江苏南通 226000
李桃	南通大学附属肿瘤医院肿瘤内科江苏南通 226001
倪静怡	南通大学附属肿瘤医院肿瘤内科江苏南通 226001
张珣磊	南通大学附属肿瘤医院肿瘤内科江苏南通 226001
张葆春	南通大学附属肿瘤医院肿瘤内科江苏南通 226001
高湘湘	南通大学附属肿瘤医院肿瘤内科江苏南通 226001

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中文摘要:

摘要目的：探讨靶向miR-592/PIK3CA的lncRNA-XIST对乳腺癌细胞裸鼠的作用机制。方法：选择2021年1月至2022年我院收集的20例乳腺癌组织及癌旁组织，对比lncRNA XIST、miR-592及PIK3CA的表达水平。培养MCF-10A、MCF-7、BT-549、MDA-MB-468、HSS578T细胞，对比不同乳腺癌细胞中lncRNA XIST、miR-592及PIK3CA的表达。建立MCF-7裸鼠移植瘤模型建立，将其分为对照组、si-NC组、si-XIST组、pcDNA-NC组、pcDNA-XIST组，对比不同组荷瘤裸鼠生存情况及各组肿瘤体积、抑瘤率。使用Western blot法检测MMP-2、MMP-9、Bax蛋白表达，之后行双荧光素酶报告基因实验。结果：乳腺癌组织中的lncRNA XIST表达水平明显较癌旁组织低，miR-592、PIK3CA表达水平明显较癌旁组织高（P<0.05）。与MCF-10A正常乳腺癌细胞相比，MCF-7乳腺癌细胞中lncRNA XIST表达最低，miR-592、PIK3CA表达最高，选择MCF-7乳腺癌细胞作为研究对象行后续研究。对照组、si-NC组、pcDNA-NC组的肿瘤体积、肿瘤重量对比无统计学意义，si-NC组、pcDNA-NC组的抑瘤率对比无统计学意义（P>0.05）；si-XIST组的肿瘤体积、肿瘤重量明显增加（P<0.05），抑瘤率为-40.04±5.24%，pcDNA-XIST组肿瘤体积、肿瘤重量明显降低（P<0.05），抑瘤率为54.29±7.89%。对照组、si-NC组、pcDNA-NC组的MMP-2、MMP-9、Bax对比无统计学意义（P>0.05）；与对照组、si-NC组、pcDNA-NC组相比，si-XIST组的MMP-2、MMP-9明显升高，Bax明显降低，pcDNA-XIST组的MMP-2、MMP-9明显降低，Bax明显升高（P<0.05）。与mimic NC相比，miR-592 mimic组中XIST-WT、PIK3CA-WT细胞中荧光活性明显降低（P<0.05），两组XIST-MUT、PIK3CA-MUT细胞中荧光活性对比无统计学意义（P>0.05）。结论：过表达XIST可能通过海绵化miR-592来调控PIK3CA表达水平，降低MMP-2、MMP-9水平，提高Bax水平，进而抑制MCF-7裸鼠皮下移植瘤的生长。

英文摘要:

ABSTRACT Objective: To investigate the mechanism of action of lncRNA-XIST targeting miR-592/PIK3CA on nude mice with breast cancer cells. Methods: The expression levels of lncRNA XIST, miR-592 and PIK3CA were compared in 20 breast cancer tissues and adjacent tissues collected in our hospital from January 2021 to 2022. MCF-10A, MCF-7, BT-549, MDA-MB-468, HSS578T cells were cultured, and the expressions of lncRNA XIST, miR-592 and PIK3CA in different breast cancer cells were compared. The transplanted tumor model of MCF-7 nude mice was established, and the mice were divided into control group, si-NC group, si-XIST group, pcDNA-NC group and pcDNA-XIST group, and the survival status, tumor volume and tumor inhibition rate of each group were compared. Western blot was used to detect the expression of MMP-2, MMP-9 and Bax proteins, and then double luciferase reporter gene assay was performed. Results: In breast cancer tissues, the expression level of lncRNA XIST was lower, and the expression levels of miR-592 and PIK3CA were higher(P<0.05). Compared with MCF-10A normal breast cancer cells, the expression of lncRNA XIST in MCF-7 breast cancer cells was the lowest, and the expression of miR-592 and PIK3CA was the highest. MCF-7 breast cancer cells were selected as the research objects for follow-up studies.There was no statistical significance in tumor volume and weight in control group, si-NC group and pcDNA-NC group, and no significance in tumor inhibition rate between si-NC group and pcDNA-NC group (P>0.05). The tumor volume and weight of the si-XIST group were increased (P<0.05), and the tumor inhibitory rate was -40.04±5.24%; the tumor volume and weight of the pcDNA-XIST group were decreased (P<0.05), and the tumor inhibitory rate was 54.29±7.89%. There was no significance in MMP-2, MMP-9 and Bax of control group, si-NC group and pcDNA-NC group(P>0.05). Compared with control group, si-NC group and pcDNA-NC group, MMP-2 and MMP-9 in si-XIST group were significantly increased, and Bax was significantly decreased, MMP-2 and MMP-9 were significantly decreased, and Bax was increased in pcDNA-XIST group(P<0.05). Compared with mimic NC, the fluorescence activity of XIST-WT and PIK3CA-WT cells in miR-592 mimic group was decreased (P<0.05), and there was no significance in the fluorescence activity of XIST-MUT and PIK3CA-MUT cells between the two groups(P>0.05). Conclusion: Overexpression of XIST may regulate PIK3CA expression level by sponging miR-592, reduce MMP-2, MMP-9 levels and increase Bax levels, and subsequently inhibit the growth of subcutaneous xenograft tumors in MCF-7 nude mice.

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