沉默REGγ基因的甲状腺癌细胞株的构建及其对p21表达的影响.docVIP

  沉默 REGγ基因的甲状腺癌细胞株的构建 及其对 p21 表达的影响# 党永岩，师凯旋，李磊，李晓涛* 5 10 15 20 25 30 35 40 （华东师范大学生命科学学院） 摘要：目的 REGγ与甲状腺癌的发生发展关系密切。沉默 REGγ基因表达有助于揭示其在 甲状腺癌发病中的生物学功能和作用机制。方法 体外培养滤泡甲状腺癌细胞株 FTC 和未分 化甲状腺癌细胞株 SW1736，用 ShRNA 干扰技术沉默这两种细胞中 REGγ基因表达，并采 用 Realtime PCR 和 Western blot 技术检测基因沉默的效果。沉默 REGγ基因表达后，Western blot 技术分析 FTC 和 SW1736 细胞中 p21 的表达情况。结果 FTC 细胞中，ShR 实验组 REG γ基因表达仅为 ShN 细胞的 12.35%，而 SW1736 细胞中，ShR 实验组 REGγ基因表达仅为 ShN 细胞的 29.49%，均有明显的抑制效果。相反，两种细胞的 ShR 实验组中 p21 的表达水 平明显增加。结论 本文使用 ShRNA 技术成功构建了沉默 REGγ基因表达的 FTC 和 SW1736 稳定细胞株，还发现沉默 REGγ基因表达后，两种不同的甲状腺癌细胞中 p21 含量均显著 增加，表明 REGγ可能通过 p21 参与甲状腺癌发生发展的调控。 关键词：REGγ，甲状腺癌，ShRNA 干扰技术，p21 中图分类号：Q7 The establishment of stable thyroid carcinoma cell lines with REGγ knockdown and its effects on p21 expression Dang Yongyan, Shi Kaixuan, Li Lei, Li Xiaotao ( School of Life Science, East china Normal University) Abstract: Objective REGγ is closely related to the pathogenesis of thyroid cancer. It is beneficial to reveal the function and mechanism of REGγ in the thyroid cancer by silencing REGγ gene expression. Methods REGγ expression was silenced by ShRNA interference in cultured follicular thyroid carcinoma (FTC) and anaplastic thyroid carcinoma cell lines (SW1736). Realtime PCR and Western blot analysis were used to detect the efficiency of ShRNA interference. Reuslts REGγ expression in ShR FTC cells were 12.35% compared with the ShN cells. In SW1736 cells, REGγ expression in ShR cells were 29.49% compared with the ShN cells. REGγ expressions were successfully inhibited in these two cell lines. P21 protein levels in both FTC cells and SW1736 cells were increased markedly. Conclusion The stable cell lines of FTC and SW1736 with REGγ knockdown were established successfully. P21 expression was increased when REGγ expression is inhibited in FTC and SW1736 cells. It seems that REGγ participates in the regulation of thyroid cancer by affecting p21 levels. Key words: REGγ, Thyroid cancer, ShRNA interference, p21 0 引言 REGγ 也称为 PSME3 (proteasome acti