Abstract: Wei Ming, Shi Guangyong, Liu Jia, Gong Yanjie, Chen Hetao, Liang Yinghong, Tu Ling Clinical Laboratory, The Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China Corresponding author: Wei Ming, Email: gushiweiming@126.com 【Abstract】 Objective To evaluate the effect of small interfering RNA（siRNA）targeting survivin gene on the of survivin and proliferation, apoptosis, migration and invasion of a human cutaneous squamous cell carcinoma cell line A431 in vitro. Methods A survivin-specific siRNA was designed and synthesized. Cultured A431 cells were divided into 3 groups to be transfected with 50.0 nmol/L liposome complexes containing survivin-specific siRNA （survivin siRNA group）, 50.0 nmol/L liposome complexes containing unrelated siRNA（negative control group） and 50.0 nmol/L prepared vesicles （blank control group）. Real-time quantitative reverse transcription-PCR （RT-PCR） and Western blot analysis were performed to determine the mRNA and protein of survivin in A431 cells, respectively. Methyl thiazolyl tetrazolium （MTT） assay was conducted to evaluate cellular proliferative activity, flow cytometry using annexin-V/propidium iodide （PI） staining to detect cell apoptosis, Transwell assay to estimate migratory and invasive activities of A431 cells, and flow cytometry to detect cell cycle changes. Results At 48 hours after transfection, the mRNA and protein of survivin both significantly differed among the survivin siRNA group, negative control group and blank control group （mRNA: 0.56 ± 0.15, 0.88 ± 0.37, 0.90 ± 0.43, F = 276.67, P < 0.001; protein: 0.59 ± 0.04, 0.86 ± 0.05, 0.91 ± 0.07, F = 243.61, P < 0.001）, the survivin siRNA group showed significantly lower mRNA and protein of survivin compared with the negative control group and blank control group （all P < 0.05）, and there were no significant differences between the negative control group and blank control group （both P > 0.05）. Repeated measures analysis of variance showed that the transfection with survivin siRNA could significantly inhibit the proliferation of A431 cells（F = 13.19, P = 0.004）, the proliferation inhibition rate was significantly higher in the survivin siRNA group than in the negative control group and blank control group （both P < 0.05）, and no significant difference was observed between the negative control group and blank control group （P > 0.05）. At 24 hours after transfection, the apoptosis rate significantly differed among the 3 groups （F = 83.97, P = 0.002）. The survivin siRNA group showed a significantly higher apoptosis rate compared with the negative control group and blank control group （both P < 0.05）, and there was no significant difference between the negative control group and blank control group （P > 0.05）. At 48 hours after transfection, the survivin siRNA group showed a significantly higher proportion of cells at G2/M phase, but lower number of migratory cells and invasive cells compared with the negative control group and blank control group （all P < 0.05）. Conclusion Survivin-specific siRNA can inhibit the of survivin gene and the proliferation of A431 cells, promote cell apoptosis, and suppress cell migration and invasion, indicating that survivin may serve as a genetic target for the treatment of cutaneous squamous cell carcinoma.

Key words: Neoplasms, squamous cell, RNA, small interfering, Cell proliferation, Apoptosis, Cell cycle, Cell movement, Survivin, A431 cells