转染微小RNA—132对骨肉瘤SaOS—2细胞生长和凋亡的作用(1)
摘要：目的 观察转染微小RNA-132 (miR-132)对体外人骨肉瘤细胞株SaOS-2生长和凋亡的影响，并初步探讨其作用机制。方法 实时荧光定量逆转录-聚合酶链反应(RT-qPCR)检测34例骨肉瘤组织及癌旁正常组织中miR-132 mRNA的表达；Cell Counting Kit-8 (CCK-8)法检测细胞增殖；细胞经DAPI染色后在荧光显微镜下观察细胞凋亡；Western blot检测体外SaOS-2细胞中Mucin-4、HER-2和p-FAK蛋白的表达。结果 miR-132 mRNA在骨肉瘤组织中的表达明显低于癌旁正常组织，差异有统计学意义(P < 0.05)；与对照组相比较，转染组细胞中miR-132 mRNA的表达明显升高，细胞増殖被明显抑制，凋亡细胞显著增加，差异均有统计学意义(P <0.05)；Mucin-4、HER-2和p-FAK蛋白在转染组细胞中的表达均下调，与对照组相比较，均有统计学差异(P<0.05)。结论 转染miR-132对体外骨肉瘤细胞有增殖抑制和凋亡诱导作用，miR-132有望成为骨肉瘤治疗的新靶点。
, 百拇医药
关键词：骨肉瘤；miR-132；凋亡
Effects of MicroRNA-132 Transfection on the Proliferation and Apoptosis in Osteosarcoma SaOS-2 Cell Line in Vitro
ZHOU Qi
(The First People Hospital of Yueyang，Yueyang 414000， Hunan，China)
Abstract：Objective To observe the biological role and underlying mechanism of miR-132 in osteosarcoma proliferation and apoptosis. Methods The expression of miR-132 mRNA in the cancer tissue and their adjacent tissues in 34 osteosarcoma patients were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The biological effects of miR-132 transfection on osteosarcoma SaOS-2 cells were assessed by CCK-8 assay and DAPI staining. Western blotting was used to detect the expression of Mucin-4， HER-2 and p-FAK in SaOS-2 cells. Results The expression level of miR-132 mRNA was found to be downregulated in osteosarcoma tissues compared with the matched adjacent tissues. After transfection， the expression of miR-132 mRNA was significantly higher than control group. The proliferation of SaOS-2 cells was inhibited significantly by miR-132 transfection. Apoptosis rate induced by the miR-132 transfection was markedly higher than that of control. The proteins of Mucin-4， HER-2 and p-FAK were decreased after miR-132 transfection. Conclusion miR-132 is downregulated in osteosarcoma tissues， transfected of miR-132 can effectively inhibit proliferation and promote apoptosis of SaOS-2 cells in vitro， miR-132 may become a new target for the regulation of gene expression in osteosarcoma.
, 百拇医药
Key words：Osteosarcoma； miR-132； Apoptosis
1引言
骨肉瘤为最常见成骨性恶性肿瘤，多发于青少年，极易复发转移，当前基因治疗逐渐成为恶性肿瘤治疗的前沿。微小RNA(microRNA， miRNAs)是一种内源性非编码小分子RNA，在组织炎症反应、细胞增殖与凋亡、组织分化以及恶性肿瘤发生和发展等多种生理过程中发挥着重要的作用[1]。其中miR-132是当前研究热点之一，miR-132不仅在结肠癌和胰腺癌等多种恶性肿瘤细胞中表达下调，同时与恶性肿瘤的转移有着密切的关系[2， 3]，表明miR-132可能作为抑制恶性肿瘤的新作用靶点。为进一步明确miR-132调控恶性肿瘤生物学行为中的作用机制，本研究运用PCR技术观察miR-132 mRNA在骨肉瘤组织和癌旁正常组织中的表达差异，并通过转染miR-132至骨肉瘤细胞，检测其对体外骨肉瘤细胞增殖和凋亡的影响，探讨miR-132作为治疗骨肉瘤新靶点的价值。
2资料与方法, http://www.100md.com(周齐)
, 百拇医药
关键词：骨肉瘤；miR-132；凋亡
Effects of MicroRNA-132 Transfection on the Proliferation and Apoptosis in Osteosarcoma SaOS-2 Cell Line in Vitro
ZHOU Qi
(The First People Hospital of Yueyang，Yueyang 414000， Hunan，China)
Abstract：Objective To observe the biological role and underlying mechanism of miR-132 in osteosarcoma proliferation and apoptosis. Methods The expression of miR-132 mRNA in the cancer tissue and their adjacent tissues in 34 osteosarcoma patients were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The biological effects of miR-132 transfection on osteosarcoma SaOS-2 cells were assessed by CCK-8 assay and DAPI staining. Western blotting was used to detect the expression of Mucin-4， HER-2 and p-FAK in SaOS-2 cells. Results The expression level of miR-132 mRNA was found to be downregulated in osteosarcoma tissues compared with the matched adjacent tissues. After transfection， the expression of miR-132 mRNA was significantly higher than control group. The proliferation of SaOS-2 cells was inhibited significantly by miR-132 transfection. Apoptosis rate induced by the miR-132 transfection was markedly higher than that of control. The proteins of Mucin-4， HER-2 and p-FAK were decreased after miR-132 transfection. Conclusion miR-132 is downregulated in osteosarcoma tissues， transfected of miR-132 can effectively inhibit proliferation and promote apoptosis of SaOS-2 cells in vitro， miR-132 may become a new target for the regulation of gene expression in osteosarcoma.
, 百拇医药
Key words：Osteosarcoma； miR-132； Apoptosis
1引言
骨肉瘤为最常见成骨性恶性肿瘤，多发于青少年，极易复发转移，当前基因治疗逐渐成为恶性肿瘤治疗的前沿。微小RNA(microRNA， miRNAs)是一种内源性非编码小分子RNA，在组织炎症反应、细胞增殖与凋亡、组织分化以及恶性肿瘤发生和发展等多种生理过程中发挥着重要的作用[1]。其中miR-132是当前研究热点之一，miR-132不仅在结肠癌和胰腺癌等多种恶性肿瘤细胞中表达下调，同时与恶性肿瘤的转移有着密切的关系[2， 3]，表明miR-132可能作为抑制恶性肿瘤的新作用靶点。为进一步明确miR-132调控恶性肿瘤生物学行为中的作用机制，本研究运用PCR技术观察miR-132 mRNA在骨肉瘤组织和癌旁正常组织中的表达差异，并通过转染miR-132至骨肉瘤细胞，检测其对体外骨肉瘤细胞增殖和凋亡的影响，探讨miR-132作为治疗骨肉瘤新靶点的价值。
2资料与方法, http://www.100md.com(周齐)