Mechanism of LncRNA XIST regulating proliferation, migration, invasion and apoptosis of breast cancer cells through miR-20a-5p/HMGA2 axis
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摘要:目的
探讨LncRNA XIST对乳腺癌细胞增殖、迁移、侵袭及凋亡的影响及机制。
方法选取正常乳腺上皮细胞MCF-10A及乳腺癌细胞系MCF-7、T47D和Bcap-37细胞，将不同物质转染至MCF-7、T47D细胞，设为空白组(无转染)、si-XIST组(转染si-XIST)、si-NC组(转染si-NC)、微小RNA-20a-5p(miR-20a-5p)组(转染miR-20a-5p mimics)及miR-NC组(转染miR-NC)。CCK-8检测细胞增殖能力; Transwell检测细胞侵袭能力; 划痕实验检测细胞迁移能力; TUNEL染色检测细胞凋亡。采用荧光素酶报告基因实验验证LncRNA XIST与miR-20a-5p及miR-20a-5p与高迁移率族蛋白A2(HMGA2)的靶向关系。采用实时荧光定量聚合酶链反应(qRT-PCR)检测LncRNA XIST、miR-20a-5p的表达水平。采用Western blot检测HMGA2的表达水平。
结果与MCF-10A细胞比较, MCF-7、T47D和Bcap-37细胞中LncRNA XIST表达水平升高, miR-20a-5p表达水平降低，差异有统计学意义(P＜0.05)。下调LncRNA XIST或上调miR-20a-5p抑制乳腺癌细胞增殖、侵袭及迁移，促进细胞凋亡(P＜0.05)。LncRNA XIST负性调控miR-20a-5p, miR-20a-5p负性调控HMGA2。
结论下调LncRNA XIST通过miR-20a-5p/HMGA2轴抑制乳腺癌细胞增殖、迁移和侵袭，促进细胞凋亡。
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关键词:
- 乳腺癌 /
- 长链非编码RNA XIST /
- 微小RNA-20a-5p /
- 高迁移率族蛋白A2
Abstract:ObjectiveTo investigate the effects and mechanisms of LncRNA XIST on proliferation, migration, invasion and apoptosis of breast cancer cells.
MethodsNormal mammary epithelial cells MCF-10A and breast cancer cell lines MCF-7, T47D and Bcap-37 cells were selected and transfected into MCF-7 and T47D cells with different substances. They were set as blank group (no transfection), si-XIST group (transfection si-XIST), si-NC group (transfection si-NC), microRNA-20a-5p group (transfection miR-20a-5p mimics) and miR-NC group (transfection miR-NC). CCK-8 was used to detect cell proliferation; Transwell was used to detect cell invasion ability; cell migration ability was detected by scratch test; apoptosis was detected by TUNEL staining. Luciferase reporter gene assay was used to verify the targeting relationships between LncRNA XIST and miR-20a-5p as well as between miR-20a-5p and high mobility group protein A2 (HMGA2). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of LncRNA XIST and miR-20a-5p. The expression level of HMGA2 was detected by Western blot.
ResultsCompared with MCF-10A cells, LncRNA XIST expression levels in MCF-7, T47D and Bcap-37 cells were significantly increased, while miR-20a-5p expression level was significantly decreased (P < 0.05). Down-regulating LncRNA XIST or up-regulating miR-20a-5p inhibited proliferation, invasion and migration of breast cancer cells, and promoted cell apoptosis (P < 0.05). LncRNA XIST negatively regulated miR-20a-5p, and miR-20a-5p negatively regulated HMGA2.
ConclusionDown-regulated LncRNA XIST inhibits proliferation, migration andinvasion of breast cancer cells and promotes cell apoptosis through miR-20a-5p/HMGA2 axis.
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图 2 沉默LncRNA XIST对乳腺癌细胞增殖、侵袭、迁移及凋亡的影响
A: LncRNA XIST表达水平; B: CCK-8结果; C: 侵袭实验结果(放大20倍); D: 迁移实验结果; E: TUNEL染色结果。
与si-NC组胞比较, *P＜0.05。![]()
图 3 上调miR-20a-5p对乳腺癌细胞增殖、侵袭、迁移及凋亡的影响
A: miR-20a-5p表达水平; B: CCK-8结果; C: 侵袭实验结果(放大20倍); D: 迁移实验结果; E: TUNEL染色结果。
与miR-NC组胞比较, *P＜0.05。![]()
图 4 LncRNA XIST与miR-20a-5p的靶向关系
A: LncRNA XIST与miR-20a-5p的结合位点; B: LncRNA XIST与miR-20a-5p相关性分析; C: 各组细胞miR-20a-5p表达水平; D: 荧光素酶活性实验。与si-NC组比较, *P＜0.05; 与miR-NC组比较, #P＜0.05。
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[1] KATSURA C, OGUNMWONYI I, KANKAM H K, et al. Breast cancer: presentation, investigation and management[J]. Br J Hosp Med (Lond), 2022, 83(2): 1-7.
[2] WINKLE M, EL-DALY S M, FABBRI M, et al. Noncoding RNA therapeutics-challenges and potential solutions[J]. Nat Rev Drug Discov, 2021, 20(8): 629-651. doi: 10.1038/s41573-021-00219-z
[3] SMOLARZ B, ZADROZNA-NOWAK A, ROMANOWICZ H. The role of LncRNA in the development of tumors, including breast cancer[J]. Int J Mol Sci, 2021, 22(16): 8427. doi: 10.3390/ijms22168427
[4] LIU Y J, ZHANG P Y, WU Q Y, et al. Long non-coding RNA NR2F1-AS1 induces breast cancer lung metastatic dormancy by regulating NR2F1 and ΔNp63[J]. Nat Commun, 2021, 12(1): 5232. doi: 10.1038/s41467-021-25552-0
[5] 张娜, 易茂林. LncRNA XIST通过海绵吸附miR-133a调控甲状腺癌细胞的增殖和凋亡[J]. 贵州医科大学学报, 2021, 46(8): 917-924. https://www.cnki.com.cn/Article/CJFDTOTAL-GYYB202108008.htm [6] 才昊, 韩亮, 李晓飞. LncRNA XIST通过调控miR-186-5p促进非小细胞肺癌的增殖和侵袭实验研究[J]. 陕西医学杂志, 2020, 49(7): 781-787, 803. doi: 10.3969/j.issn.1000-7377.2020.07.004 [7] YAN Z Z, LI J, GUO J, et al. LncRNA XIST sponges microRNA-448 to promote malignant behaviors of colorectal cancer cells via regulating GRHL2[J]. Funct Integr Genomics, 2022, 22(5): 977-988. doi: 10.1007/s10142-022-00873-5
[8] 胡炜, 邵安静, 李春霞, 等. 沉默miR-20a-5p的肝癌细胞恶性生物学行为及Zbed3、Axin蛋白表达情况观察[J]. 山东医药, 2022, 62(11): 28-32. https://www.cnki.com.cn/Article/CJFDTOTAL-SDYY202211007.htm [9] LI J, YE D L, SHEN P, et al. MiR-20a-5p induced WTX deficiency promotes gastric cancer progressions through regulating PI3K/AKT signaling pathway[J]. J Exp Clin Cancer Res, 2020, 39(1): 212. doi: 10.1186/s13046-020-01718-4
[10] XU J X, FANG X J, LONG L Y, et al. HMGA2 promotes breast cancer metastasis by modulating Hippo-YAP signaling pathway[J]. Cancer Biol Ther, 2021, 22(1): 5-11. doi: 10.1080/15384047.2020.1832429
[11] ZHAO W Y, GENG D H, LI S Q, et al. LncRNA HOTAIR influences cell growth, migration, invasion, and apoptosis via the miR-20a-5p/HMGA2 axis in breast cancer[J]. Cancer Med, 2018, 7(3): 842-855. doi: 10.1002/cam4.1353
[12] LI P, WANG L H, LI P F, et al. Silencing lncRNA XIST exhibits antiproliferative and proapoptotic effects on gastric cancer cells by up-regulating microRNA-132 and down-regulating PXN[J]. Aging (Albany NY), 2020, 13(10): 14469-14481.
[13] WANG H R, LI H M, YU Y K, et al. Long non-coding RNA XIST promotes the progression of esophageal squamous cell carcinoma through sponging miR-129-5p and upregulating CCND1 expression[J]. Cell Cycle, 2021, 20(1): 39-53. doi: 10.1080/15384101.2020.1856497
[14] ZHANG M, WANG F, XIANG Z, et al. LncRNA XIST promotes chemoresistance of breast cancer cells to doxorubicin by sponging miR-200c-3p to upregulate ANLN[J]. Clin Exp Pharmacol Physiol, 2020, 47(8): 1464-1472. doi: 10.1111/1440-1681.13307
[15] 郭会, 方艳君, 李翠娥, 等. 过/降表达miR-20a-5p的骨肉瘤细胞增殖、侵袭能力及Bax、Bcl-2、E2F5蛋白表达观察[J]. 山东医药, 2022, 62(11): 42-46. https://www.cnki.com.cn/Article/CJFDTOTAL-SDYY202211010.htm [16] HE Y, MA H, WANG J, et al. MiR-20a-5p inhibits endometrial cancer progression by targeting Janus kinase 1[J]. Oncol Lett, 2021, 21(5): 427. doi: 10.3892/ol.2021.12688
[17] YANG H B, CHEN Z, LIU Z M. MiR-20a-5p negatively regulates NR4A3 to promote metastasis in bladder cancer[J]. J Oncol, 2021, 2021: 1377989.
[18] BAI X D, HAN G H, LIU Y, et al. MiRNA-20a-5p promotes the growth of triple-negative breast cancer cells through targeting RUNX3[J]. Biomed Pharmacother, 2018, 103: 1482-1489. doi: 10.1016/j.biopha.2018.04.165
[19] 李会芳, 侯永强, 王明军, 等. 乳腺癌组织FOXP3、HMGA2、RASAL2的表达及与临床病理特征和预后的关系[J]. 现代生物医学进展, 2021, 21(20): 3959-3964. https://www.cnki.com.cn/Article/CJFDTOTAL-SWCX202120032.htm [20] 李晨阳, 张学红. MiR-154靶向HMGA2基因在人子宫内膜癌中的表达及其对癌细胞恶性生物学行为的影响[J]. 兰州大学学报: 医学版, 2022, 48(1): 21-27. https://www.cnki.com.cn/Article/CJFDTOTAL-LZYX202201006.htm
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