张文军, 郑立春, 柴连海, 张晓明, 夏安庆, 胡耀杰. Wip1在甲状腺癌细胞中表达的临床及生物学意义[J]. 中国肿瘤临床, 2014, 41(21): 1348-1352. DOI: 10.3969/j.issn.1000-8179.20140291
引用本文: 张文军, 郑立春, 柴连海, 张晓明, 夏安庆, 胡耀杰. Wip1在甲状腺癌细胞中表达的临床及生物学意义[J]. 中国肿瘤临床, 2014, 41(21): 1348-1352. DOI: 10.3969/j.issn.1000-8179.20140291
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ZHANG Wenjun, ZHENG Lichun, ZHANG Xiaoming, XIA Anqing, HU Yaojie, CHAI Lianhai. Expression and clinical significance of Wip1 in thyroid carcinoma and biological effect of siRNA targeting Wip1 on its cell line[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2014, 41(21): 1348-1352. DOI: 10.3969/j.issn.1000-8179.20140291
Citation: ZHANG Wenjun, ZHENG Lichun, ZHANG Xiaoming, XIA Anqing, HU Yaojie, CHAI Lianhai. Expression and clinical significance of Wip1 in thyroid carcinoma and biological effect of siRNA targeting Wip1 on its cell line[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2014, 41(21): 1348-1352. DOI: 10.3969/j.issn.1000-8179.20140291
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Wip1在甲状腺癌细胞中表达的临床及生物学意义

Expression and clinical significance of Wip1 in thyroid carcinoma and biological effect of siRNA targeting Wip1 on its cell line

    摘要
  • 摘要:
      目的  探讨野生型p53诱导的磷酸酶1(wild-type p53-induced phosphatase 1, Wip1)在甲状腺癌中的表达意义及其导入靶向Wip1的siRNA对甲状腺乳头状癌K1细胞的生物学影响。
      方法  采用免疫组织化学方法、逆转录聚合酶链反应(reverse transcription polymerase chain reaction, RT-PCR)检测73例甲状腺癌组织及距其癌组织边缘5 cm以上的镜下未见浸润的正常组织中蛋白、mRNA的表达情况。通过脂质体介导将Wip1基因siRNA瞬时转染甲状腺乳头状癌K1细胞, 采用RT-PCR及Western blot检测转染后K1细胞内Wip1基因的表达水平, MTT比色法及流式细胞术(FCM)观察细胞增殖、凋亡、周期变化情况。
      结果  Wip1蛋白在甲状腺癌组织及正常甲状腺组织中的表达阳性率分别为80.8%、9.6%, 较正常甲状腺组织明显升高(χ2=47.036, P < 0.05)。Wip1 mRNA在甲状腺癌组织及正常甲状腺组织中的表达相对量分别为0.665±0.046、0.225±0.039, 较正常甲状腺组织明显升高(t=12.637, P < 0.05)。Wip1蛋白、mRNA表达与甲状腺癌患者性别、年龄及肿瘤大小无关(P>0.05), 而与淋巴结转移、肿瘤临床分期及肿瘤分化有关(P < 0.05)。RT-PCR及Western blot证实, siRNA沉默的K1细胞中Wip1 mRNA、蛋白的表达水平较未转染组明显降低(t=17.039, t=14.637, P < 0.05)。MTT、细胞凋亡及周期结果表明, siRNA沉默的K1细胞其增殖能力明显减弱、细胞凋亡明显增加, G0+G1期细胞数比例升高, 而G2+M期和S期细胞数比例降低(P < 0.05)。
      结论  甲状腺癌组织中Wip1蛋白和mRNA的表达均升高, 且与甲状腺癌淋巴结转移、肿瘤临床分期以及肿瘤分化有关, Wip1参与甲状腺癌细胞增殖、凋亡、周期这一生物学过程。

     

    Abstract:
      Objective  To investigate the expression and clinical significance of wild-type p53-induced phosphatase 1(Wip1) in thyroid carcinoma and biological effect of siRNA-targeting Wip1 on the thyroid carcinoma cell line.
      Methods  Immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) were performed to detect the expression of Wip1 in 73 specimens of thyroid carcinoma tissues and normal thyroid tissues (5 cm away from the margin of thyroid carcinoma), respectively.Wip1 siRNA was transiently transfected into the papillary thyroid carcinoma cell by using a liposome-mediated method and then detected by RT-PCR and Western blot.Methyl thiazolyl tetrazolium (MTT) assay and flow cytometry (FCM) were also conducted to observe cell proliferation, cell apoptosis, and cell cycle.
      Results  The positive rates of Wip1 protein were 80.8% in thyroid carcinoma tissues and 9.6% in the normal tissues (χ2=47.036, P < 0.05).The relative mRNA contents of Wip1 were 0.665 ± 0.046 and 0.225 ± 0.039 in carcinoma and normal tissues, respectively; these results significantly differed between the two types (t=12.637, P < 0.05).Significant correlation was not observed between Wip1 expression and other factors, such as patient's gender, age, and tumor size (P>0.05).However, significant correlations among Wip1 expression, lymph node metastasis, clinical stages and tumor differentiation (P < 0.05) were observed.RT-PCR and Western blot results showed that K1 cell-transfected Wip1 siRNA exhibited a relatively lower expression than normal cells (t=17.039, t= 14.637, P < 0.05).MTT assay results showed that the K1 cells transfected with Wip1 siRNA showed a lower survival fraction, higher cell apoptosis, higher percentage of G0/G1 phases, and lower cell concentration in G2/M and S phases (P < 0.05).
      Conclusion  Wip1 protein and mRNA were increased in thyroid carcinoma and are correlated with lymph node metastasis, clinical stages and tumor differentiation.Wip1 may be involved in proliferation, apoptosis, and cycle of thyroid cancer cells.

     

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