"Biomarker" Research Laboratory, Institute of Fundamental Biology and Medicine, Kazan Federal University, 420008 Kazan, Russia.
Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA 19111, USA.
Molecules. 2022 Feb 14;27(4):1289. doi: 10.3390/molecules27041289.
The chemoresistance of tumor cells is one of the most urgent challenges in modern oncology and in pancreatic cancer, in which this problem is the most prominent. Therefore, the identification of new chemosensitizing co-targets may be a path toward increasing chemotherapy efficacy. In this work, we performed high-performance in vitro knockout CRISPR/Cas9 screening to find potential regulators of the sensitivity of pancreatic cancer. For this purpose, MIA PaCa-2 cells transduced with two sgRNA libraries ("cell cycle/nuclear proteins genes" and "genome-wide") were screened by oxaliplatin and cisplatin. In total, 173 candidate genes were identified as potential regulators of pancreatic cancer cell sensitivity to oxaliplatin and/or cisplatin; among these, 25 genes have previously been reported, while 148 genes were identified for the first time as potential platinum drug sensitivity regulators. We found seven candidate genes involved in pancreatic cancer cell sensitivity to both cisplatin and oxaliplatin. Gene ontology enrichment analysis reveals the enrichment of single-stranded DNA binding, damaged DNA binding pathways, and four associated with NADH dehydrogenase activity. Further investigation and validation of the obtained results by in vitro, in vivo, and bioinformatics approaches, as well as literature analysis, will help to identify novel pancreatic cancer platinum sensitivity regulators.
肿瘤细胞的化疗耐药性是现代肿瘤学中最紧迫的挑战之一，而在胰腺癌中，这个问题最为突出。因此，鉴定新的化疗增敏共靶标可能是提高化疗疗效的途径。在这项工作中，我们进行了高通量体外 CRISPR/Cas9 敲除筛选，以寻找潜在的调节胰腺癌细胞敏感性的药物。为此，用两种 sgRNA 文库（“细胞周期/核蛋白基因”和“全基因组”）转导的 MIA PaCa-2 细胞用奥沙利铂和顺铂进行筛选。总共鉴定出 173 个候选基因，这些候选基因可能是调节胰腺癌细胞对奥沙利铂和顺铂敏感性的潜在调节剂；其中，25 个基因以前曾被报道过，而 148 个基因则首次被鉴定为潜在的铂类药物敏感性调节剂。我们发现了七个候选基因，这些基因参与了胰腺癌细胞对顺铂和奥沙利铂的敏感性。基因本体富集分析揭示了单链 DNA 结合、损伤 DNA 结合途径以及与 NADH 脱氢酶活性相关的四个途径的富集。通过体外、体内和生物信息学方法以及文献分析对获得的结果进行进一步的研究和验证，将有助于鉴定新的胰腺癌细胞铂类药物敏感性调节剂。
