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。
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.
关键词
甲状腺癌； /
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miR-133-3p； /
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细胞增殖； /
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SOD； /
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细胞凋亡
Key words
Thyroid cancer; /
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miR-133-3p; /
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Cell proliferation; /
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SOD; /
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Cell apoptosis
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基金
河南省医学科技攻关计划项目（2018020748）
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