Tan Lulu, Chen Zhihua
Department of Radiation Oncology, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, Hainan, China.
Department of Digestive Medicine, Haikou People's Hospital, Haikou, Hainan, China.
J Oncol. 2022 Oct 14;2022:8071343. doi: 10.1155/2022/8071343. eCollection 2022.
This study was to investigate whether miR-193a-5p and ZFP57 are involved in the radioresistance of pancreatic cancer and to explore its working mechanism. Pancreatic cancer tissues were harvested from patients who achieved CR (complete remission) and PR (partial remission) and those who achieved PD (progressive disease) and SD (stable disease). The mRNA and protein expressions of ZFP57 and miR-193a-5p were determined by RT-qPCR and WB (Western blot), respectively. For experiments, the parental BxPC-3 cell line was irradiated by X-ray at a total dose of 40 Gy to induce the irradiation-resistant subtype BxPC-3-RR. ZFP57 was downregulated in radioresistant pancreatic cancer cells. The results of dual-luciferase reporter gene assay, RNA pull-down assay, RT-qPCR, and WB confirmed that miR-193a-5p targeted ZFP57 and inhibited ZFP57 expression. The MTT assay and the colony formation assay showed that the radioresistant pancreatic cancer cells had higher viability and survival fraction. The results of WB indicated that in the radioresistant pancreatic cancer cells, the cyclin D1, Bax, CDk4, cleaved caspase-3, Bcl-2, and -H2AX proteins were upregulated to varying degrees. The results of the nude mouse experiment were consistent with those of experiments. According to the cell transfection and salvage experiments, miR-193a-5p down regulated ZFP57 after radiotherapy. As a result, the Wnt pathway was activated, which further induced radioresistance of pancreatic cancer cells. Our experiments showed that the miR-193a-5p/ZFP57/Wnt pathway mediated the radioresistance of pancreatic cancer cells, providing novel clues for the treatment of pancreatic cancer.
本研究旨在探讨miR-193a-5p和ZFP57是否参与胰腺癌的放射抗性，并探索其作用机制。从达到完全缓解（CR）和部分缓解（PR）的患者以及疾病进展（PD）和疾病稳定（SD）的患者中获取胰腺癌组织。分别通过逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法（WB）检测ZFP57和miR-193a-5p的mRNA和蛋白质表达。在实验中，亲代BxPC-3细胞系接受40 Gy的X射线照射以诱导抗辐射亚型BxPC-3-RR。ZFP57在抗辐射的胰腺癌细胞中表达下调。双荧光素酶报告基因检测、RNA下拉检测、RT-qPCR和WB的结果证实miR-193a-5p靶向ZFP57并抑制ZFP57表达。MTT检测和集落形成检测表明，抗辐射的胰腺癌细胞具有更高的活力和存活分数。WB结果表明，在抗辐射的胰腺癌细胞中，细胞周期蛋白D1、Bax、细胞周期蛋白依赖性激酶4、裂解的半胱天冬酶-3、Bcl-2和γ-H2AX蛋白均有不同程度的上调。裸鼠实验结果与细胞实验结果一致。根据细胞转染和挽救实验，放疗后miR-193a-5p下调ZFP57。结果，Wnt信号通路被激活，进一步诱导胰腺癌细胞的放射抗性。我们的实验表明，miR-193a-5p/ZFP57/Wnt信号通路介导了胰腺癌细胞的放射抗性，为胰腺癌的治疗提供了新的线索。
