RNA干扰SALL4基因对儿童睾丸卵黄囊瘤细胞增殖和凋亡的影响

doi:10.3969/j.issn.2095-9400.2019.12.006

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摘要目的：研究干扰SALL4基因对儿童睾丸卵黄囊瘤细胞的体外增殖、凋亡的影响。方法：通过构建SALL4干扰质粒载体pGPU6/GFP/Neo-SALL4 shRNA，转染儿童睾丸卵黄囊瘤细胞，并在mRNA和蛋白水平验证干扰效果，通过CCK-8、流式细胞术来研究其对儿童睾丸卵黄囊瘤细胞体外增殖、凋亡的影响。结果：成功构建载体并干扰SALL4表达后，荧光定量PCR结果示干扰组mRNA水平（24.0%±4.0%）较阴性对照组（93.0%± 43.0%）显著下降（P＜0.05），蛋白相对表达量（0.39±0.10）也较阴性对照组（1.02±0.09）显著降低（P＜0.05），儿童睾丸卵黄囊瘤细胞的体外增殖活力显著降低，细胞凋亡率显著上升（P＜0.05）。结论：RNA靶向干扰SALL4表达可抑制儿童睾丸卵黄囊瘤细胞增殖能力并诱导其凋亡，提示SALL4基因可能是儿童睾丸卵黄囊瘤一个潜在的治疗靶点。

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关键词 ： RNA干扰, SALL4, 儿童, 睾丸卵黄囊瘤

Abstract：Objective: To study the effect of RNA interference gene SALL4 on the proliferation and apaptosis of testicular yolk sac tumor cells in children. Methods: The pGPU6/GFP/neo-sall4 shRNA was constructed as a vector to be transfected into pediatric testicular yolk sac tumor cells and interfere in SALL4. The interfering effect was verified by detecting mRNA and protein levels. CCK-8 and flow cytometry were employed to study the inhibition effect of interfering SALL4 in pediatric testicular yolk sac tumor cells. Results: After successfully transfecting the vector into pediatric testicular yolk sac tumor cells, the mRNA expression magnitudes of SALL4 interfered group was 24.0%±4.0%, as compared with Negative group (93.0%±43.0%), with lower levels of mRNA expression (P<0.05). And the protein expressions of SALL4 interfered group (0.39±0.10) had a lower protein expression than Negative group (1.02±0.09). Besides, the proliferation was significantly reduced, while the apoptosis rate was significantly increased in vitro (P<0.05). Conclusion: Targeting SALL4 RNA could inhibit the proliferation and induce apoptosis of pediatric testicular yolk sac tumor cells, suggesting that SALL4 may be a potential therapeutic target for pediatric testicular yolk sac tumor.

Key words： RNA interference SALL4 children testicular yolk sac tumor

收稿日期: 2019-07-15

基金资助:温州市科技计划项目（Y20140491）。

通讯作者: 陈肖鸣，主任医师，教授，Email：cxm@wzmc.edu.cn。

作者简介: 吕敏（1987-），男，浙江绍兴人，住院医师，硕士。

链接本文:

https://xb.wmu.edu.cn/CN/10.3969/j.issn.2095-9400.2019.12.006 或 https://xb.wmu.edu.cn/CN/Y2019/V49/I12/885

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