Javadrashid Darya, Mohammadzadeh Reza, Baghbanzadeh Amir, Safaee Sahar, Amini Mohammad, Lotfi Ziba, Baghbani Elham, Khaze Shahgoli Vahid, Baradaran Behzad
Immunology Research Center, Tabriz University of Medical Sciences, Iran.
Faculty of Basic Sciences, Department of Biology, University of Maragheh, Iran.
EXCLI J. 2021 Jan 21;20:160-173. doi: 10.17179/excli2020-2900. eCollection 2021.
Despite the recent advances in the treatment of other cancers, the 5-year survival rate of pancreatic cancer remains under 9 %. Chemotherapy and surgical resection are the most common therapy methods. The regulatory role of microRNAs in different types of cancer has given them therapeutic importance. miR-612 has been downregulated in colorectal, bladder, liver, and some other types of cancer and could be considered a tumor-suppressor miRNA. 5-FU is one of the most common chemotherapeutic agents used in pancreatic cancer treatment, which is used in multiple drug regimens and combinatorial therapy methods. The aim of this study is the evaluation of miR-612 restoration in the PANC-1 cell line and using the tumor-suppressive effect of it in combination with 5-FU on cell growth and migration. MiR-612 mimic was transfected to PANC-1 cells through electroporation. Following the transfection, expression levels of miR-612 and BAX, BCL-2, Caspase-3, MMP9, and PD-L1 genes were measured by qRT-PCR. MTT assay was used to determine the cytotoxicity of miR-612 and 5-FU on PANC-1 cell viability. To confirm MTT results and to evaluate the quantitative effect of apoptosis induction flow cytometry test was used and in order to confirm apoptosis test results and cell cycle arrest evaluation DAPI staining and cell, cycle tests were conducted, respectively. Finally, to assess the inhibitory effect of miR-612 in combination with 5-FU on migration and growth wound healing and colony formation assays were used, respectively. Results demonstrated that miR-612 alongside 5-FU has an important role in the inhibition of migration and growth and also apoptosis induction in PANC-1 cells and could be considered as a supporting agent of chemotherapy and a novel therapeutic modality in pancreatic cancer treatment.
尽管近期在其他癌症的治疗方面取得了进展，但胰腺癌的5年生存率仍低于9%。化疗和手术切除是最常见的治疗方法。微小RNA在不同类型癌症中的调控作用使其具有治疗重要性。miR-612在结直肠癌、膀胱癌、肝癌和其他一些类型的癌症中表达下调，可被视为一种肿瘤抑制性微小RNA。5-氟尿嘧啶是胰腺癌治疗中最常用的化疗药物之一，用于多种联合用药方案和联合治疗方法中。本研究的目的是评估miR-612在PANC-1细胞系中的恢复情况，并利用其肿瘤抑制作用与5-氟尿嘧啶联合作用于细胞生长和迁移。通过电穿孔将miR-612模拟物转染到PANC-1细胞中。转染后，通过qRT-PCR检测miR-612以及BAX、BCL-2、Caspase-3、MMP9和PD-L1基因的表达水平。采用MTT法测定miR-612和5-氟尿嘧啶对PANC-1细胞活力的细胞毒性。为了证实MTT结果并评估凋亡诱导的定量效果，使用了流式细胞术检测，为了证实凋亡检测结果和评估细胞周期阻滞，分别进行了DAPI染色和细胞周期检测。最后，分别采用伤口愈合试验和集落形成试验评估miR-612与5-氟尿嘧啶联合对迁移和生长的抑制作用。结果表明，miR-612与5-氟尿嘧啶一起在抑制PANC-1细胞的迁移和生长以及诱导凋亡方面具有重要作用，可被视为化疗的辅助药物和胰腺癌治疗的一种新型治疗方式。
