Department of Bioengineering, Integral University, Lucknow, India.
Department of Biotechnology, Noida Institute of Engireering and Technology, Greater Noida, India.
Anticancer Agents Med Chem. 2019;19(16):2019-2033. doi: 10.2174/1871520619666190725122400.
The aberrant alteration in Jab1 signalosome (COP9 Signalosome Complex Subunit 5) has been proven to be associated with the progression of several carcinomas. However the specific role and mechanism of action of Jab1 signalosome in carcinogenesis of gall bladder cancer (GBC) are poorly understood.
The main objective of our study was to elucidate the role and mechanism of Jab1 signalosome in gall bladder cancer by employing siRNA.
Jab1 overexpression was identified in gall bladder cancer tissue sample. The role of Jab1-siRNA approach in cell growth inhibition and apoptotic induction was then examined by RT-PCR, Western Blotting, MTT, ROS, Hoechst and FITC/Annexin-V staining.
In the current study, we have shown that overexpression of Jab1 stimulated the proliferation of GBC cells; whereas downregulation of Jab1 by using Jab1-siRNA approach resulted incell growth inhibition and apoptotic induction. Furthermore, we found that downregulation of Jab1 induces cell cycle arrest at G1 phase and upregulated the expression of p27, p53 and Bax gene. Moreover, Jab1-siRNA induces apoptosis by enhancing ROS generation and caspase-3 activation. In addition, combined treatment with Jab1-siRNA and gemicitabine demonstrated an enhanced decline in cell proliferation which further suggested increased efficacy of gemcitabine at a very lower dose (5μM) in combination with Jab1-siRNA.
In conclusion, our study strongly suggests that targeting Jab1 signalosome could be a promising therapeutic target for the treatment of gall bladder cancer.
Jab1 信号体（COP9 信号体复合物亚基 5）的异常改变已被证明与几种癌的进展有关。然而，Jab1 信号体在胆囊癌（GBC）发生中的具体作用和作用机制仍知之甚少。
本研究的主要目的是通过 siRNA 阐明 Jab1 信号体在胆囊癌中的作用和机制。
鉴定了胆囊癌组织样本中 Jab1 的过表达。然后通过 RT-PCR、Western Blotting、MTT、ROS、Hoechst 和 FITC/Annexin-V 染色检测 Jab1-siRNA 方法在细胞生长抑制和诱导凋亡中的作用。
在本研究中，我们表明 Jab1 的过表达刺激了 GBC 细胞的增殖；而使用 Jab1-siRNA 方法下调 Jab1 导致细胞生长抑制和凋亡诱导。此外，我们发现下调 Jab1 诱导细胞周期停滞在 G1 期，并上调 p27、p53 和 Bax 基因的表达。此外，Jab1-siRNA 通过增强 ROS 生成和 caspase-3 激活诱导细胞凋亡。此外，Jab1-siRNA 和吉西他滨联合治疗显示细胞增殖下降幅度更大，这进一步表明在联合使用 Jab1-siRNA 时，吉西他滨在非常低的剂量（5μM）下的疗效增强。
总之，我们的研究强烈表明，靶向 Jab1 信号体可能是治疗胆囊癌的一种有前途的治疗靶点。
