miR-140-5p对非小细胞肺癌细胞增殖及迁移的抑制作用

miR-140-5p Inhibits Proliferation and Migration of Non-small Cell Lung Cancer Cells

目的研究miR-140-5p对非小细胞肺癌细胞增殖和迁移能力的影响。方法 qRT-PCR测定miR-140-5p在非小细胞肺癌细胞系HCC827、H1650、H1975、H1299、A549及正常肺上皮细胞系BEAS-2B中的表达。将A549细胞系分成3组:miR-140-5p过表达组(miR-140-5p组)、miR-140-5p阴性对照组(miR-NC组)及空白对照组(Mock组),经LipofectamineTM2000分别转染过表达序列pcDNA6.2-GW/miR-140-5p mimics、阴性对照序列pcDNA6.2-GW/microRNA scramble及空白对照质粒。CCK-8实验测定细胞增殖能力,细胞划痕实验测定迁移能力。蛋白印迹测定PIN-1及PDGFRa蛋白表达。结果 miR-140-5p在非小细胞肺癌细胞系HCC827(P<0.05)、H1650(P<0.05)、H1975(P<0.01)、H1299(P<0.01)及A549(P<0.01)中表达量均低于人正常肺上皮细胞系BEAS-2B。转染后72h及96h,CCK-8实验结果显示miR-140-5p组vs.miR-NC组的A540nm值分别为(0.53±0.06)vs.(0.83±0.08)(P<0.05)、(0.68±0.06)vs.(1.17±0.10)(P<0.01)。miR-140-5p组划痕愈合率低于miR-NC组,(33.71±2.39)%vs.(57.94±5.48)%,P<0.05。miR-140-5p组PIN-1、PDGFRa蛋白相对表达量低于miR-NC组(P<0.05)。结论 miR-140-5p在非小细胞肺癌细胞系中低表达,过表达miR-140-5p可抑制细胞增殖和迁移,可能与PIN-1、PDGFRa蛋白表达下调有关。

Objective To study the effects of miR-140-5p on proliferation and migration of non-small cell lung cancer cell line.Methods The expression of miR-140-5p in non-small cell lung cancer cell lines,HCC827,H1650,H1975,H1299and A549and normal lung cell line,BEAS-2B,was quantified by qRT-PCR.The A549cell line was divided into three groups,miR-140-5p over-expression group(miR-140-5p group),miR-140-5p negative control group(miR-NC group)and blank control group(Mock group),which was transfected with pcDNA6.2-GW/miR-140-5p mimics,pcDNA6.2-GW/microRNA scramble and blank plasmid,respectively.CCK-8assay and wound scratch assay was used to measure the proliferation and migration ability,respectively.The expression level of PIN-1and PDGFRa protein was measured by Western blotting.Results miR-140-5p was significantly down-regulated in non-small cell lung cancer cell lines HCC827(P<0.05),H1650(P<0.05),H1975(P<0.01),H1299(P<0.01)and A549(P<0.01),compared with normal lung cell line,BEAS-2B.The A540nm value of miR-140-5p group and miR-NC group was(0.53±0.06)vs.(0.83±0.08)(P<0.05),(0.68±0.06)vs.(1.17±0.10)(P<0.01),respectively,after transfection for72and96h.Scar healing rate of miR-140-5p group was significantly lower than that of miR-NC group[(33.71±2.39)%vs.(57.94±5.48)%,P<0.05].The relative protein expression level of PIN-1and PDGFRa was significantly lower in the miR-140-5p group than in the miR-NC group(P<0.05).Conclusion miR-140-5p expression is down-regulated in non-small cell lung cancer cell lines.Over-expression of miR-140-5p can inhibit cell proliferation and migration,which may be associated with down-regulation of PIN-1and PDGFRa protein.