Profiling the Panc1 pancreatic cancer cell line following Gemcitabine treatment using single cell RNA sequencing

Pancreatic ductal adenocarcinoma (PDAC) often presents at late clinical stages, and most patients are managed solely through palliative chemotherapy. With no approved treatment modalities for patients who progress on broad-spectrum chemotherapy, we set to identify druggable targets to prevent or reverse resistance to the first line anti-neoplastic Gemcitabine. In our first experiment, we used the well-established Panc1 cell line as an in vitro model of PDAC. Panc1 cells were incubated with a tolerable dose of Gemcitabine in vitro, and examined alterations in gene expression via single cell RNA sequencing. In our subsequent studies, we incubated Panc1 cells with increasing doses of Gemcitabine for several passages, until viable in approximately 10x the known IC50 value. These cells were designated Panc1-GR. Based on our observations in the prior experiment, Panc1-GR cells were compared to those treated with wither the Calmodulin inhibitor W-7, Calcium chelator BAPTA-AM, or the calcium channel blocker Amlodipine. Through these efforts, we hope to better understand the mechanisms of Gemcitabine resistance in PDAC, as well as introduce new therapeutic strategies to reverse drug resistant phenotypes in the clinic. Examining gemcitabine resistance in pancreatic cancer using single cell RNA-seq

胰腺导管腺癌（Pancreatic ductal adenocarcinoma, PDAC）往往在临床晚期才被确诊，多数患者仅能接受姑息性化疗。针对广谱化疗后病情进展的患者，目前尚无获批的治疗方案，本研究旨在识别可成药靶点，以预防或逆转一线抗肿瘤药物吉西他滨（Gemcitabine）的耐药性。
在首次实验中，我们采用已被广泛使用的Panc1细胞系作为PDAC的体外模型，将Panc1细胞以耐受剂量的吉西他滨进行体外孵育，并通过单细胞RNA测序（single cell RNA sequencing）分析基因表达的变化。
在后续研究中，我们使用逐步递增剂量的吉西他滨对Panc1细胞进行持续多代孵育培养，直至细胞在约为已知半抑制浓度（IC50）10倍的药物浓度下仍可存活，将该耐药细胞系命名为Panc1-GR。
基于前期实验的观察结果，我们将Panc1-GR细胞分别与经钙调蛋白抑制剂W-7、钙螯合剂BAPTA-AM或钙通道阻滞剂氨氯地平（Amlodipine）处理的细胞进行对照。
通过本研究的一系列实验，我们期望更深入地阐明PDAC中吉西他滨耐药的分子机制，并为临床逆转肿瘤耐药表型提供全新的治疗策略。
基于单细胞RNA测序探究胰腺癌吉西他滨耐药性