RPL35A通過介導HSPA8泛素化促進膽管癌的進展

2024年2月，中華醫學院**醫院肝膽外科、鞍山市中心醫院普通外科南51號、****醫學中心普通外科高級科中國人民解放軍總醫院（1Hepatobiliary Surgery Department, First Hospital of China Medical University, ****55, Nanjingbei street, 110001 Shenyang, Liaoning Province, P. R. China. 2Department of General Surgery, Anshan Central Hospital, No.51, South Zhonghua Road, Tiedong District, 114008 Anshan, Liaoning Province, China. 3Senior Department of General Surgery, the First Medical Center of Chinese PLA General Hospital, No.28, Fuxing Road, Haidian District,

100039 Beijing, China）Ruizhao Qi3 and Jialin Zhang1研究團隊在《Biology Direct》上發表論文：

“RPL35A promotes the progression of cholangiocarcinoma by mediating HSPA8 ubiquitination”

“RPL35A通過介導HSPA8泛素化促進膽管癌的進展”

Abstract：

Background Cholangiocarcinoma (CCA) is a biliary epithelial malignant tumor with an increasing incidence worldwide. Therefore, further understanding of the molecular mechanisms of CCA progression is required to identify new therapeutic targets. Methods The expression of RPL35A in CCA and para-carcinoma tissues was detected by immunohistochemical

staining. IP-MS combined with Co-IP identified downstream proteins regulated by RPL35A. Western blot and Co-IP of CHX or MG-132 treated CCA cells were used to verify the regulation of HSPA8 protein by RPL35A. Cell experiments and subcutaneous tumorigenesis experiments in nude mice were performed to evaluate the effects of RPL35A and HSPA8 on the proliferation, apoptosis, cell cycle, migration of CCA cells and tumor growth in vivo.

Results RPL35A was significantly upregulated in CCA tissues and cells. RPL35A knockdown inhibited the proliferation

and migration of HCCC-9810 and HUCCT1 cells, induced apoptosis, and arrested the cell cycle in G1 phase. HSPA8 was a downstream protein of RPL35A and overexpressed in CCA. RPL35A knockdown impaired HSPA8 protein stability and

increased HSPA8 protein ubiquitination levels. RPL35A overexpression promoted CCA cell proliferation and migration.

HSPA8 knockdown inhibited CCA cell proliferation and migration, and reversed the promoting effect of RPL35A.

Furthermore, RPL35A promoted tumor growth in vivo. In contrast, HSPA8 knockdown suppressed tumor growth, while was able to restore the effects of RPL35A overexpression.

Conclusion RPL35A was upregulated in CCA tissues and promoted the progression of CCA by mediating HSPA8 ubiquitination.

摘要：

背景

膽管癌(CCA)是一種膽道上皮惡性腫瘤，在世界范圍內發病率不斷上升。因此，需要進一步了解CCA進展的分子機制，以確定新的治療靶點。

方法

免疫組化染色檢測RPL35A在CCA和癌旁組織中的表達。IP-MS聯合Co-IP鑒定了RPL35A調控的下游蛋白。采用Western blot和Co-IP對CHX或MG-132處理的CCA細胞進行檢測，驗證RPL35A對HSPA8蛋白的調控作用。通過細胞實驗和裸鼠皮下腫瘤發生實驗，在體內評價RPL35A和HSPA8對CCA細胞增殖、凋亡、細胞周期、遷移和腫瘤生長的影響。

結果

RPL35A在CCA組織和細胞中顯著上調。RPL35A敲低抑制hcc -9810和HUCCT1細胞的增殖和遷移，誘導細胞凋亡，使細胞周期停留在G1期。HSPA8是RPL35A的下游蛋白，在CCA中過表達。RPL35A敲低會損害HSPA8蛋白的穩定性，增加HSPA8蛋白的泛素化水平。RPL35A過表達促進CCA細胞增殖和遷移。HSPA8敲低抑制CCA細胞增殖和遷移，逆轉RPL35A的促進作用。此外，RPL35A在體內促進腫瘤生長。相反，HSPA8敲低抑制腫瘤生長，同時能夠恢復RPL35A過表達的效果。

結論

RPL35A在CCA組織中表達上調，通過介導HSPA8泛素化促進CCA的進展。

該論文中，4種膽管癌細胞系(hcc -9810、HUCCT1、QBC939、RBE細胞)的體外培養是使用Ausbian特級胎牛血清完成的。欲了解或購買Ausbian特級胎牛血清可以聯系北京締一生物400-166-8600.