miR-302b过表达对人舌癌细胞株TSCCa增殖、迁移和侵袭的影响

Effects of miR 302b overexpression on proliferation,migration and invasion of human tongue cancer cell line TSCCa

目的研究miR-302b过表达对人舌癌细胞株TSCCa增殖、迁移和侵袭的影响及其潜在的分子机制。方法用miR-302b-NC和miR-302b mimic转染舌癌细胞TSCCa,命名为miR-302b-NC组和miR-302b mimic组,以未转染的细胞为对照,转染48 h后,采用流式细胞仪检测转染效率,实时荧光定量PCR(qRT-PCR)检测TSCCa细胞中miR-302b的表达情况,分别采用MTT实验、划痕实验和Transwell小室检测TSCCa细胞增殖、迁移和侵袭能力,采用蛋白免疫印迹(Western blot)检测TSCCa细胞中增殖细胞核抗原(PCNA)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)表达情况。结果流式细胞仪检测结果显示转染效率达85%以上;转染后,miR-302b mimic组人舌癌细胞TSCCa中miR-302b的表达水平明显高于对照组和miR-302b-NC组(P <0. 05); miR-302b mimic组人舌癌细胞TSCCa增殖率、迁移能力和侵袭能力显著低于对照组和miR-302b-NC组(P <0. 05); miR-302b mimic组人舌癌细胞TSCCa中PCNA、MMP-2、MMP-9蛋白的表达水平明显低于对照组和miR-302b-NC组(P <0. 05)。结论 miR-302b过表达能够抑制人舌癌细胞TSCCa增殖、迁移和侵袭,推测这一作用是通过调控PCNA、MMP-2、MMP-9的表达水平实现的,为舌癌的靶向治疗提供一定的理论依据。

Objective To investigate the effect of miR 302b overexpression on proliferation, migration and invasion of human tongue cancer cell line TSCCa and its potential molecular mechanism. MethodsThe tongue cancer cell line TSCCa was transfected with miR 302b NC and miR 302b mimic, named as miR 302b NC group and miR 302b mimic group, and the non transfected cells were regarded as control group. At 48 hours after transfection, the transfection efficiency was detected by flow cytometry, and the miR 302b expression in TSCCa cells was detected by quantitative real time fluorescence quantification PCR (qRT PCR). MTT assay, scratch test and transwell chamber test were used to detect the proliferation, migration and invasion ability of TSCCa cells respectively. The expression levels of proliferating cell nuclear antigen (PCNA), matrix metalloproteinase 2 (MMP-2) and MMP-9 in TSCCa cells were detected by Western Blot. ResultsFlow cytometry results showed that transfection efficiency was above 85%. After transfection, the miR 302b expression level of human tongue cancer cell TSCCa in miR 302b mimic group was significantly higher than that in the control group and miR 302b NC group ( P < 0.05 ). The proliferation, migration and invasion abilities of human tongue cancer cell TSCCa in the miR 302b mimic group were significantly lower than those in control group and miR 302b NC group ( P <0.05). The expression levels of PCNA, MMP 2 and MMP 9 of human tongue cancer cell TSCCa in miR 302b mimic group were significantly lower than those in control group and miR 302b NC group ( P <0.05). ConclusionThe overexpression of miR 302b can inhibit the proliferation, migration and invasion of human tongue cancer cell TSCCa, it is speculated that the effects are realized by regulating the expression levels of PCNA, MMP-2 and MMP-9, which provides a theoretical basis for targeted therapy of tongue cancer.