Zhou Min, Gao Yang, Wang Min, Guo Xingjun, Li Xu, Zhu Feng, Xu Simiao, Qin Renyi
Department of Biliary-Pancreatic Surgery, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Department of Endocrinology, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 430030, Wuhan, China.
J Cancer. 2021 May 3;12(12):3726-3740. doi: 10.7150/jca.48418. eCollection 2021.
Cancer stem cells (CSCs) initiate and maintain tumorigenesis due to their unique pluripotency. However, pancreatic stem cell gene signatures are not completely revealed yet. Here, we isolated pancreatic cancer stem cells (P-CSCs) and exploited their distinct genome-wide mRNA and miRNA expression profiles using microarrays. CD24 CD44 ESA cells were isolated from two pancreatic xenograft cells by the flow cytometry and identified the stem cell-like properties by the tumor formation, self-renew and chemoresistance. Microarrays and qRT-PCR were used to exploit their distinct Genome-wide mRNA and miRNA expression profiles. The function and candidate target genes of key microRNA were detected after Ectopic restoration in the pancreatic cancer cell lines MIA Paca-2 (CSC) and BxPC-3 (CSC). In this study, we isolated P-CSCs from two xenografts cells. Genome-wide profiling experiments showed 479 genes and 15 microRNAs specifically expressed in the P-CSCs, including genes involved in TGF-β and p53 signaling pathways and particularly miR-146b-3p as the most significantly downregulated miRNA. We confirmed miR-146b-3p as a downregulated signature in pancreatic cancer tissues and cell line MIA Paca-2 (CSC) cells. Ectopic restoration of miR-146b-3p expression with pre-miR reduced cell proliferation, induced apoptosis, increased G1 phase and reduced S phase in cell cycle in MIA Paca-2 (CSC), but not in BxPC-3 (CSC). Re-expression of miR-146b-3p with lentivirus significantly inhibited tumorigenicity in MIA Paca-2, but slightly in BxPC-3. Furthermore, we demonstrated that miR-146b-3p directly targeted MAP3K10 and might activate Hedgehog pathway as well through DYRK2 and GLI2. These results suggest that P-CSCs have distinct gene expression profiles. MiR-146b-3p inhibits proliferation and induced apoptosis in P-CSCs high cells lines by targeting MAP3K10. Targeting P-CSCs specific genes may provide novel strategies for therapeutic purposes.
癌症干细胞（CSCs）因其独特的多能性启动并维持肿瘤发生。然而，胰腺干细胞基因特征尚未完全揭示。在此，我们分离出胰腺癌干细胞（P-CSCs），并使用微阵列技术研究其独特的全基因组mRNA和miRNA表达谱。通过流式细胞术从两种胰腺异种移植细胞中分离出CD24 CD44 ESA细胞，并通过肿瘤形成、自我更新和化学抗性鉴定其干细胞样特性。使用微阵列和qRT-PCR研究其独特的全基因组mRNA和miRNA表达谱。在胰腺癌细胞系MIA Paca-2（CSC）和BxPC-3（CSC）中进行异位恢复后，检测关键microRNA的功能和候选靶基因。在本研究中，我们从两种异种移植细胞中分离出P-CSCs。全基因组分析实验显示，479个基因和15个microRNA在P-CSCs中特异性表达，包括参与TGF-β和p53信号通路的基因，特别是miR-146b-3p是下调最显著的miRNA。我们证实miR-146b-3p是胰腺癌组织和细胞系MIA Paca-2（CSC）细胞中的下调特征。用pre-miR异位恢复miR-146b-3p表达可降低MIA Paca-2（CSC）细胞的增殖，诱导凋亡，增加细胞周期中的G1期并减少S期，但对BxPC-3（CSC）细胞无此作用。用慢病毒重新表达miR-146b-3p可显著抑制MIA Paca-2细胞的致瘤性，但对BxPC-3细胞的抑制作用较弱。此外，我们证明miR-146b-3p直接靶向MAP3K10，并可能通过DYRK2和GLI2激活Hedgehog通路。这些结果表明P-CSCs具有独特的基因表达谱。MiR-146b-3p通过靶向MAP3K10抑制P-CSCs高表达细胞系中的增殖并诱导凋亡。靶向P-CSCs特异性基因可能为治疗目的提供新策略。
