Up-regulation of small GTPases, RhoA and RhoC, is associated with tumor progression in ovarian carcinoma
Up-regulation of small GTPases, RhoA and RhoC, is associated with tumor progression in ovarian carcinoma
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DOI：
10.1097/01.lab.0000073128.16098.31
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发表时间：
2003-06-01
影响因子：
5
通讯作者：
Konishi, I
中科院分区：
文献类型：
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作者：
Horiuchi, A;Imai, T;Konishi, I
为了阐明小GTP酶Rho在卵巢癌生物学行为中的作用，我们首先使用RT-PCR和实时RT-PCR检测了良性、交界性和恶性卵巢肿瘤中RhoA、RhoB和RhoC的mRNA表达。采用Western blotting和免疫组织化学方法分析RhoA蛋白的表达和定位。最后，我们研究了Rho的上调是否增强了卵巢癌细胞的体外侵袭力。Rho家族基因的mRNA水平分析显示，癌中RhoA和RhoC的水平显著高于良性肿瘤（RhoA，p = 0.0035; RhoC，p = 0.0006）。浆液性癌RhoA和RhoC mRNA表达水平均显著高于其他组织学类型。关于国际妇产科联合会分期分类，RhoA和RhoC mRNA水平在III+IV期肿瘤中均显著高于I+II期肿瘤（RhoA，p = 0.0200; RhoC，p = 0.0057）。此外，分析配对的原发性和播散性病变表明，RhoA和RhoC mRNA的表达在转移性肿瘤中显著高于原发性肿瘤。RhoA蛋白表达水平的检测表明，在晚期卵巢癌中，尤其是浆液性卵巢癌中，RhoA的表达也增加。因此，我们推测Rho GTP酶的上调在卵巢癌的进展中起重要作用。使用卵巢癌细胞系SKOV 3的基质胶侵袭测定显示，用溶血磷脂酸处理后的上调和活化与癌细胞的增强的侵袭相关。这种侵袭性的增加被C3抑制，C3是Rho的特异性抑制剂。这些发现表明Rho GTPases的上调在卵巢癌的肿瘤进展中是重要的，并且Rho家族蛋白可能是癌症治疗的分子靶点。
To clarify the role of small GTPases Rho in the biologic behavior of ovarian carcinoma, we first examined the mRNA expression of RhoA, RhoB, and RhoC in benign, borderline, and malignant ovarian tumors using RT-PCR and real-time RT-PCR. The expression and localization of RhoA protein were also analyzed by Western blotting and immunohistochemistry. Finally, we examined whether up-regulation of Rho enhances the invasiveness of ovarian cancer cells in vitro. Analysis of mRNA levels of the Rho family genes revealed that levels of both RhoA and RhoC were significantly higher in carcinomas than in benign tumors (RhoA, p = 0.0035; RhoC, p = 0.0006). According to histologic subtype, both RhoA and RhoC mRNA levels in serous carcinomas were significantly higher than those in other histologic types. With regard to the International Federation of Gynecological and Obstetrics stage classification, both of RhoA and RhoC mRNA levels were significantly higher in tumors of Stages III+IV than in those of Stages I+II (RhoA, p = 0.0200; RhoC, p = 0.0057). In addition, analysis of matched pairs of primary and disseminated lesions demonstrated that expression of both RhoA and RhoC mRNA was significantly higher in metastatic than in primary tumors. Examination of the protein level showed that expression of RhoA was also increased in advanced ovarian carcinomas, especially those of serous histology. Accordingly, we hypothesized that up-regulation of Rho GTPases plays an important role in the progression of ovarian carcinoma. Matrigel invasion assay using the ovarian cancer cell line, SKOV3, showed that up-regulation and activation after treatment with lysophosphatidic acid was associated with enhanced invasion of the cancer cells. This increase in invasiveness was suppressed by the addition of C3, a specific inhibitor of Rho. These findings suggest that up-regulation of Rho GTPases is important in the tumor progression of ovarian carcinoma and that Rho family proteins could be a molecular target in cancer therapy.