Fhit基因在子宫内膜癌增殖、侵袭中的作用和临床意义

Role and clinical significance of Fhit gene in the proliferation and invasion of endometrial carcinoma

目的:探讨Fhit基因在子宫内膜癌中的表达情况,并通过RNA激活上调Fhit表达后观察其对子宫内膜癌细胞增殖和侵袭、迁移的影响。方法:收集135例子宫内膜癌组织和40例正常子宫内膜组织,使用免疫组织化学和Western blotting检测Fhit基因的表达情况并分析Fhit表达与临床病理特征的联系。构建Fhit-saRNA表达载体,转染至子宫内膜癌ISK细胞株,建立上调Fhit基因表达的子宫内膜癌细胞株。Western blotting和RT-PCR验证转染效果。通过CCK-8和Transwell试验分析转染前后增殖、迁移、侵袭能力的差异。结果:免疫组织化学和Western blotting结果均显示在子宫内膜癌组织中Fhit基因表达与正常子宫内膜组织相比明显降低甚至缺失(P<0.01);CCK-8结果显示Fhit基因表达上调后子宫内膜癌细胞生长明显减慢,与未转染组比较差异有统计学意义(P<0.01);Transwell试验结果显示Fhit基因表达上调后子宫内膜癌细胞迁移和侵袭能力显著减弱,空白对照组的穿膜细胞数显著高于转染组(P<0.01)。结论:相比于正常子宫内膜组织,Fhit基因在子宫内膜癌组织中表达较低且与临床病理特征相关。在子宫内膜癌细胞株中通过saRNA上调Fhit后,其增殖、迁移、侵袭能力受到明显抑制。

Objective:To explore the expression level of Fhit gene in endometrial carcinoma,and observe its effects on the proliferation,invasion and migration of endometrial carcinoma cells through the RNA activation up-regulating the expresson of Fhit.Methods:The expression levels of Fhit protein in 135 endometrial carcinoma tissues and 40 normal endometrial tissues were detected using immunohistochemistry and Western blotting.The correlation between Fhit expression and clinicopathological features was analyzed.The Fhit-saRNA expression vector was constructed,and transfected into endometrial carcinoma ISK cell line,and the endometrial carcinoma cell line with Fhit up-regulating expression was established.The transfection effects were verified using Western blotting and RT-PCR.The differences of the proliferation,migration and invasion abilities between before and after transfection were analyzed using CCK-8 and Transwell assays.Results:The results of immunohistochemistry and Western blotting showed that the expression levels of Fhit in endometrial carcinoma tissue significantly reduced or even missed compared with normal endometrial tissue(P<0.01).The results of CCK-8 showed that the growth of endometrial carcinoma significantly slowed down after the up-regulating of Fhit expression,and the difference of which between transfection group and non-transfection group was statistically significant(P<0.01).The results of Transwell assay showed that the migration and invasion abilities of endometrial carcinoma cells after the level of Fhit up-regulating significantly weakened,and the number of transmembrane cells in blank control group were significantly higher than that in transfection group(P<0.01).Conclusions:Compared with normal endometrial tissues,the expression level of Fhit in endometrial carcinoma tissues is lower,and related to the clinicopathological features.After up-regulating the Fhit expression by RNA activation in endometrial carcinoma cell line,the abilities of proliferation,migration and invasion of cells are significantly inhibited.