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目的:探讨靶向生长分化因子15(growth differentiation factor 15,GDF15)基因siRNA对人肺腺癌SPCA1细胞增殖和迁移的影响。方法:根据基因数据库(Gen Bank)设计并合成3条人GDF15基因siRNA(GDF15-siRNA161、GDF15-siRNA290和GDF15-siRNA860)和阴性对照NC-siRNA序列,采用qPCR法测定转染不同GDF15-siRNA的SPCA1细胞中GDF15 mRNA表达水平,筛选有效抑制GDF15基因表达的siRNA。CCK-8法和划痕实验分别检测转染有效GDF15-siRNA和NC-siRNA的SPCA1细胞增殖和迁移能力。结果:3条GDF15-siRNA对SPCA1细胞GDF15 mRNA表达均有明显抑制(P<0.01),其中GDF15-siRNA161抑制最为明显。和转染NC-siRNA的SPCA1细胞相比,转染GDF15-siRNA161的SPCA1细胞增殖明显减慢[(0.46±0.03)vs(0.52±0.01),P<0.01],转染GDF15-siRNA161的SPCA1细胞迁移速度明显低于转染NC-siRNA的SPCA1细胞(P<0.01)。结论:有效沉默人肺腺癌SPCA1细胞GDF15基因表达能抑制肺腺癌细胞的增殖和迁移,GDF15可能成为人肺腺癌基因治疗的候选靶点。
Objective: To investigate the effect of siRNA targeting GDF15 gene on the proliferation and migration of human lung adenocarcinoma SPCA1 cells. METHODS: Three human GDF15 siRNAs (GDF15-siRNA161, GDF15-siRNA290 and GDF15-siRNA860) and negative control NC-siRNA sequences were designed and synthesized according to GenBank. QPCR method was used to determine the expression of GDF15- SPCA1 cells GDF15 mRNA expression levels, screening effectively inhibit GDF15 gene expression of siRNA. The proliferation and migration of SPCA1 cells transfected with effective GDF15-siRNA and NC-siRNA were detected by CCK-8 assay and scratch assay respectively. Results: Three GDF15-siRNAs significantly inhibited GDF15 mRNA expression in SPCA1 cells (P <0.01), of which GDF15-siRNA161 inhibited the most obviously. The proliferation of SPCA1 cells transfected with GDF15-siRNA161 was significantly slower than that of SPCA1 cells transfected with NC-siRNA [(0.46 ± 0.03) vs (0.52 ± 0.01), P <0.01]. SPCA1 cells transfected with GDF15-siRNA161 The migration speed was significantly lower than that of SPCA1 cells transfected with NC-siRNA (P <0.01). Conclusion: GDF15 gene silencing effectively inhibits the proliferation and migration of lung adenocarcinoma cells in human lung adenocarcinoma SPCA1 cells. GDF15 may be a potential target for gene therapy of human lung adenocarcinoma.