miR-133-3p对甲状腺癌SW579细胞生物学行为和移植瘤生长的影响

doi:10.13418/j.issn.1001-165x.2023.1.10

摘要/Abstract

摘要： 目的探究miR-133-3p对甲状腺癌SW579细胞和体内移植瘤生长的影响。方法甲状腺癌SW579细胞分为对照组，miR-NC组，miR-133-3p mimic组，按分组转染miR-NC和miR-133-3p mimic。qRT-PCR检测甲状腺癌组织和癌旁组织及转染后细胞中miR-133-3p的表达，克隆形成；EDU染色检测细胞增殖；流式检测细胞凋亡和线粒体膜电位变化；Western blot检测Survivin，caspase-3蛋白表达；试剂盒检测细胞上清液中SOD和MDA含量。裸鼠皮下注射转染miR-133-3p mimic或miR-NC的SW579细胞，qRT-PCR检测瘤组织中miR-133-3p表达量，记录瘤组织体积，TUNEL检测瘤组织凋亡，免疫组化检测瘤组织中Survivin和caspase-3的表达。结果甲状腺癌组织中miR-133-3p的表达降低。体外细胞实验结果显示，与对照组相比，miR-133-3p组miR-133-3p表达量增加，细胞克隆形成率、EDU阳性细胞率和Survivin蛋白表达降低，细胞凋亡率和caspase-3的蛋白表达升高，线粒体膜电位降低，细胞上清液中SOD含量降低，MDA含量增加，均P<0.05。体内实验结果显示，与对照组相比，miR-133-3p组移植瘤体积明显减小，瘤组织中miR-133-3p表达量、TUNEL阳性细胞率和caspae-3阳性细胞率均增加，Survivin阳性细胞率减少，均P<0.05。结论 miR-133-3p能够抑制甲状腺癌SW579细胞增殖，诱导细胞凋亡及线粒体氧化损伤，进而抑制裸鼠皮下移植瘤生长，有望成为甲状腺癌基因治疗的靶点。

关键词: 甲状腺癌； , , miR-133-3p； , , 细胞增殖； , , SOD； , , 细胞凋亡

Abstract: Objective To explore the effect of miR-133-3p on the growth of thyroid cancer SW579 cells and transplanted tumors in vivo. Methods Thyroid cancer SW579 cells were divided into a control group, a miR-NC group, a miR-133-3p mimic group, and transfected with miR-NC and miR-133-3p mimic according to groups. qRT-PCR was used to detect the expression of miR-133-3p in thyroid cancer tissues and adjacent tissues and transfected cells, and clone formation. EDU staining were employed to detect cell proliferation, flow cytometry was performed to detect cell apoptosis and mitochondrial membrane potential changes. The protein expression of Survivin and caspase-3 were detected by Western blot. The kits were commended to detect the content of SOD and MDA in the cell supernatant. SW579 cells transfected with miR-133-3p mimic or miR-NC were subcutaneously injected into nude mice, and the expression of miR-133-3p was detected by qRT-PCR, the volume of tumor tissue was observed and recorded. TUNEL was employed to detected tumor tissue apoptosis, and immunohistochemistry was used to detect the expression of Survivin and caspase-3 in tumor tissue. Results The expression of miR-133-3p in thyroid cancer tissues was decreased. Compared with the control group, the expression of miR-133-3p in the miR-133-3p group was significantly increased (P<0.05). The rate of cell colony formation, the rate of EDU positive cells and the expression of Survivin protein were significantly reduced (P<0.05). The apoptosis rate and the expression of caspase-3 were significantly increased (P<0.05). The mitochondrial membrane potential and the SOD content were significantly decreased, and the MDA content was increased (P<0.05) in the cell supernatant. Compared with the control group, the size of transplanted tumors in the miR-133-3p group was significantly reduced (P<0.05), the expression of miR-133-3p, the rate of TUNEL positive cells and the rate of caspae3 positive cells were significantly increased in tumor tissues (P<0.05), and the rate of Survivin-positive cells was significantly reduced (P<0.05). Conclusions miR-133-3p can inhibit the proliferation of thyroid cancer SW579 cells, induce apoptosis and mitochondrial oxidative damage, thereby inhibiting the growth of subcutaneous transplanted tumors in nude mice, which is expected to become a target for gene therapy of thyroid cancer.

Key words: Thyroid cancer; , , miR-133-3p; , , Cell proliferation; , , SOD; , , Cell apoptosis

中图分类号:

R736.1

段方方, 侯小霞, 周寒丽, 陈露, 王留晏, 赵晓丽, 张玉洁, 孔天东. miR-133-3p对甲状腺癌SW579细胞生物学行为和移植瘤生长的影响[J]. 中国临床解剖学杂志, 2023, 41(1): 51-57.

Duan Fangfang, Hou Xiaoxia, Zhou Hanli, Chen Lu, Wang Liuyan, Zhao Xiaoli, Zhang Yujie, Kong Tiandong. Effects of miR-133-3p on the biological behavior of thyroid cancer SW579 cells and the growth of transplanted tumors[J]. Chinese Journal of Clinical Anatomy, 2023, 41(1): 51-57.

参考文献 20

[1]	Qiu K, Xie Q, Jiang S, et al. miR-98-5p promotes apoptosis and inhibits migration and cell growth in papillary thyroid carcinoma through Bax/Caspase-3 by HMGA2[J]. J Clin Lab Anal, 2020, 34(2): e23044. DOI: 10.1002/jcla.23044.
[2]	吴闽, 侯慧珍, 司廷林. 过表达miR-138抑制TCF-4对甲状腺癌细胞CAL-62增殖, 凋亡和运动的调节作用[J]. 中国临床解剖学杂志, 2019, 37(4): 425-430. DOI: 10.13418/j.issn.1001-165x.2019.04.013.
[3]	Asghar MY, Lassila T, Paatero I, et al. Stromal interaction molecule 1 (STIM1) knock down attenuates invasion and proliferation and enhances the expression of thyroid-specific proteins in human follicular thyroid cancer cells[J]. Cell Mol Life Sci, 2021, 78(15): 5827-5846. DOI: 10.1007/s00018-021-03880-0.
[4]	Vigneri R, Malandrino P, Russo M. Is thyroid cancer increasing in incidence and aggressiveness [J]? J Clin Endocrinol Metab, 2020, 105(7): dgaa223. DOI: 10.1210/clinem/dgaa223.
[5]	Roth MY, Witt RL, Steward DL. Molecular testing for thyroid nodules: review and current state[J]. Cancer, 2018, 124(5): 888-898. DOI: 10.1002/cncr.30708.
[6]	Liu H, Lei C, He Q, et al. Nuclear functions of mammalian MicroRNAs in gene regulation, immunity and cancer[J]. Mol Cancer, 2018, 17(1): 64-78. DOI: 10.1186/s12943-018-0765-5.
[7]	黄天舒, 裴丽鹏. miR-135a靶向PTEN促进宫颈癌细胞增殖、凋亡和转移能力的机制研究[J]. 解放军医药杂志, 2021, 33(1): 20-26. DOI: 10.3969/j.issn.2095-140X.2021.01.005.
[8]	Curtale G. MiRNAs at the crossroads between innate immunity and cancer: focus on macrophages[J]. Cells, 2018, 7(2): 12-37. DOI: 10.3390/cells7020012.
[9]	潘庆春, 喻望博, 李四军. miR-133调控FGFR1蛋白对喉癌细胞迁移和侵袭的影响[J]. 解放军医药杂志, 2018, 30(12): 36-40. DOI: 10.3969/j.issn.2095-140X.2018.12.009.
[10]	Li TF, Liu J, Fu SJ. The interaction of long non-coding RNA MIAT and miR-133 play a role in the proliferation and metastasis of pancreatic carcinoma[J]. Biomed Pharmacother, 2018, 104: 145-150. DOI: 10.1016/j.biopha.2018.05.043.
[11]	Fang L, Kong D, Xu W. MicroRNA-625-3p promotes the proliferation, migration and invasion of thyroid cancer cells by up-regulating astrocyte elevated gene 1[J]. Biomed Pharmacother, 2018, 102: 203-211. DOI: 10.1016/j.biopha.2018.03.043.
[12]	Zhou Y, Wu D, Tao J, et al. MicroRNA-133 inhibits cell proliferation, migration and invasion by targeting epidermal growth factor receptor and its downstream effector proteins in bladder cancer[J]. Scand J Urol, 2013, 47(5): 423-432. DOI: 10.3109/00365599.2012.748821.
[13]	Liu S, Chen J, Zhang T, et al. MicroRNA-133 inhibits the growth and metastasis of the human lung cancer cells by targeting epidermal growth factor receptor[J]. J BUON, 2019, 24(3): 929-935. PMID: 31424644.
[14]	Liu Y, Han L, Bai Y, et al. Down-regulation of microRNA-133 predicts poor overall survival and regulates the growth and invasive abilities in glioma[J]. Artif Cells Nanomed Biotechnol, 2018, 46(1): 206-210. DOI: 10.1080/21691401.2017.1304551.
[15]	Liu G, Li YI, Gao X. Overexpression of microRNA-133b sensitizes non-small cell lung cancer cells to irradiation through the inhibition of glycolysis[J]. Oncol Lett, 2016, 11(4): 2903-2908. DOI: 10.3892/ol.2016.4316.
[16]	姚锐, 郑胡忠, 吴丽群, 等. 甘草酸通过线粒体膜去极化诱导HPV18阳性的人宫颈癌HeLa细胞凋亡的作用研究[J]. 中国现代应用药学, 2020,37(22): 2727-2733. DOI: 10.13748/j.cnki.issn1007-7693. 2020. 22. 007.
[17]	陈杰, 佟玲, 朱亦峰. 龙胆苦苷对卵巢癌细胞株SKOV3氧化应激损伤和细胞凋亡的影响及机制研究[J]. 医学分子生物学杂志, 2019, 16(2): 126-131. DOI: 10.3870/j.issn.1672-8009.2019.02.005.
[18]	陈炜, 陈志刚, 彭君臣, 等. 异鼠李素调控卵巢癌细胞生长、凋亡和氧化应激反应的机制[J]. 医学分子生物学杂志, 2020, 17(5): 353-358. DOI: 10.3870/j.issn.1672-8009.2020.05.002.
[19]	张丹妮, 王洪新. 肌肽对高糖诱导的大鼠H9C2心肌细胞损伤的保护作用及其机制[J]. 中国临床解剖学杂志, 2021, 39(4): 437-442. DOI: 10.13418/j.issn.1001-165x.2021.04.014.
[20]	Hassanshahi J, Mirzahosseini-Pourranjbar A, Hajializadeh Z, et al. Anticancer and cytotoxic effects of troxerutin on hela cell line: An in-vitro model of cervical cancer[J]. Mol Biol Rep, 2020, 47(8): 6135-6142. DOI: 10.1007/s11033-020-05694-y.