Su Ying, Zhao An, Cheng Guoping, Xu Jingjing, Ji Enming, Sun Wenyong
Department of Pathology, Zhejiang Cancer Hospital, Hangzhou, Zhejiang, China.
Cancer Research Institute, Zhejiang Cancer Hospital, Hangzhou, Zhejiang, China.
Cancer Biomark. 2017 Jul 4;19(3):289-296. doi: 10.3233/CBM-160449.
Renal cell carcinoma (RCC) is the highest mortality rate of the genitourinary cancers, and the treatment options are very limited. Thus, identification of molecular mechanisms underlying RCC tumorigenesis, is critical for identifying biomarkers for RCC diagnosis and prognosis.
To validate whether the IGF-I/JAK2-STAT3/miR-21 signaling pathway is associated with human RCC cell growth.
qRT-PCR and Western blotting were used to detect the mRNA and protein expression levels, respectively. The MTT assay was performed to determine cell survival rate. The Annexin V-FITC/PI apoptosis detection kit was used to detect cell apoptosis. We employed RCC tissues and cell lines (A498; ACHN; Caki-1; Caki-2 and 786-O) in the study. IGF-I, and its inhibitor (NT-157) were administrated to detect the effects of IGF-I on the expression of miR-21 and p-JAK2. JAK2 inhibitor (AG490), and si-STAT3 were used to detect the effects of JAK2/STAT3 signaling pathway on the expression of miR-21.
In our study, we firstly showed that the expression levels of IGF-I and miR-21 were up-regulated in RCC tissues and cell lines. After exogenous IGF-I treatment, the expression levels of miR-21, p-IGF-IR and p-JAK2 were significantly increased, whereas NT-157 treatment showed the reversed results. Further study indicated that JAK2 inhibitor or si-STAT3 significantly reversed the IGF-I-induced miR-21 expression level. Finally, we found that IGF-I treatment significantly prompted human RCC cell survival and inhibited cell apoptosis, and NT-157 treatment showed the reversed results.
The IGF-I/JAK2-STAT3/miR-21 signaling pathway may be associated with human RCC cell growth.
肾细胞癌（RCC）是泌尿生殖系统癌症中死亡率最高的，且治疗选择非常有限。因此，确定RCC肿瘤发生的分子机制对于识别RCC诊断和预后的生物标志物至关重要。
验证IGF-I/JAK2-STAT3/miR-21信号通路是否与人类RCC细胞生长相关。
分别使用qRT-PCR和蛋白质印迹法检测mRNA和蛋白质表达水平。采用MTT法测定细胞存活率。使用Annexin V-FITC/PI凋亡检测试剂盒检测细胞凋亡。本研究采用RCC组织和细胞系（A498；ACHN；Caki-1；Caki-2和786-O）。给予IGF-I及其抑制剂（NT-157）以检测IGF-I对miR-21和p-JAK2表达的影响。使用JAK2抑制剂（AG490）和si-STAT3检测JAK2/STAT3信号通路对miR-21表达的影响。
在我们的研究中，我们首先表明IGF-I和miR-21的表达水平在RCC组织和细胞系中上调。外源性IGF-I处理后，miR-21、p-IGF-IR和p-JAK2的表达水平显著增加，而NT-157处理则显示相反的结果。进一步研究表明，JAK2抑制剂或si-STAT3显著逆转了IGF-I诱导的miR-21表达水平。最后，我们发现IGF-I处理显著促进了人类RCC细胞存活并抑制了细胞凋亡，而NT-157处理则显示相反的结果。
IGF-I/JAK2-STAT3/miR-21信号通路可能与人类RCC细胞生长相关。
