Department of Hepatobiliary and Pancreatic Surgery, The Second Hospital of Jilin University, Changchun 130041, P.R. China.
Department of Radiation Oncology, The Second Hospital of Jilin University, Changchun 130041, P.R. China.
Biosci Rep. 2020 Sep 30;40(9). doi: 10.1042/BSR20194509.
Some microRNAs (miRs or miRNAs) have been reported to function as tumor suppressors in gallbladder cancer (GBC). However, the specific effect of miR-205-5p on GBC remains unclear. The objective of the present study was to unravel the effects of miR-205-5p on the drug resistance in GBC. For this purpose, the expression of miR-205-5p and protein kinase C ϵ (PRKCE) was quantified in the peripheral blood sample harvested from GBC patients and healthy volunteers. Then the relationship between miR-205-5p and PRKCE was validated. After isolating the GBC stem cells, ectopic expression and depletion experiments were conducted to analyze the effect of miR-205-5p and PRKCE on cell proliferation, drug resistance, apoptosis, and colony formation rate as well as the expression of apoptotic factors (Bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), and cleaved caspase 3). Finally, the mouse xenograft model of GBC was established to verify the function of miR-205-5p in vivo. Intriguingly, our results manifested that miR-205-5p was down-regulated, while PRKCE was up-regulated in peripheral blood samples and stem cells of patients with GBC. Moreover, miR-205-5p targeted PRKCE and negatively regulated its expression. The overexpression of miR-205-5p or silencing of PRKCE inhibited the drug resistance, proliferation, and colony formation rate while promoting apoptosis of GBC stem cells. Additionally, the overexpression of miR-205-5p attenuated drug resistance to gemcitabine but promoted the gemcitabine-induced cell apoptosis by inhibiting the PRKCE in vivo. Overall, an intimate correlation between miR-205-5p and PRKCE is a key determinant of drug resistance of GBC stem cells, thus, suggesting a novel miR-205-5p-based clinical intervention target for GBC patients.
一些 microRNAs(miRs 或 miRNAs)已被报道在胆囊癌(GBC)中作为肿瘤抑制因子发挥作用。然而,miR-205-5p 对 GBC 的具体影响尚不清楚。本研究的目的是揭示 miR-205-5p 对 GBC 耐药性的影响。为此,定量检测了 GBC 患者和健康志愿者外周血样本中 miR-205-5p 和蛋白激酶 C ε(PRKCE)的表达。然后验证了 miR-205-5p 和 PRKCE 之间的关系。分离 GBC 干细胞后,进行异位表达和耗尽实验,以分析 miR-205-5p 和 PRKCE 对细胞增殖、耐药性、凋亡和集落形成率以及凋亡因子(B 细胞淋巴瘤 2 相关 X 蛋白(Bax)、B 细胞淋巴瘤 2(Bcl-2)和裂解半胱天冬酶 3)表达的影响。最后,建立 GBC 小鼠异种移植模型以验证 miR-205-5p 在体内的功能。有趣的是,我们的结果表明,miR-205-5p 在 GBC 患者的外周血样本和干细胞中下调,而 PRKCE 上调。此外,miR-205-5p 靶向 PRKCE 并负调控其表达。miR-205-5p 的过表达或 PRKCE 的沉默抑制了 GBC 干细胞的耐药性、增殖和集落形成率,同时促进了其凋亡。此外,miR-205-5p 的过表达在体内抑制了吉西他滨耐药,但通过抑制 PRKCE 促进了吉西他滨诱导的细胞凋亡。总体而言,miR-205-5p 和 PRKCE 之间的密切相关性是 GBC 干细胞耐药性的关键决定因素,因此,为 GBC 患者提供了一种新的基于 miR-205-5p 的临床干预靶点。