JuBei Oral Liquid Induces Mitochondria-Mediated Apoptosis in NSCLC Cells

Zhenzhen Pan; Qiufang Chen; Xiulan Zheng; Kai Wang; Yalei Duan; Kang Xiao; Zhirong Jia; Xuansheng Ding

doi:10.2147/OTT.S254464

JuBei Oral Liquid Induces Mitochondria-Mediated Apoptosis in NSCLC Cells

Onco Targets Ther. 2020 Jul 31:13:7585-7598. doi: 10.2147/OTT.S254464. eCollection 2020.

Authors

Zhenzhen Pan^#¹, Qiufang Chen^#², Xiulan Zheng¹, Kai Wang¹, Yalei Duan¹, Kang Xiao¹, Zhirong Jia¹, Xuansheng Ding¹

Affiliations

¹ Department of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing 211198, People's Republic of China.
² Department of Science and Education, Women and Children's Hospital, School of Medicine, Xiamen University, Xiamen 361003, People's Republic of China.

^# Contributed equally.

Abstract

Background: Although gefitinib brings about tremendous advances in the treatment of non-small cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) mutations, most of patients become incurable due to drug resistance. JuBei oral liquid (JB) has been widely used to treat pneumonia in clinic. Components of JB were reported to induce apoptosis in NSCLC, which indicated that JB could be a potential antitumor agent for NSCLC patients. In this study, we investigated the effect of JB on gefitinib-sensitive PC-9 and gefitinib-resistant PC-9/GR, H1975 cells as well as its underlying molecular mechanisms.

Methods: PC-9, PC-9/GR and H1975 cells were treated with JB, LY294002, SCH772984, gefitinib alone or in combination. Then, cell viability, colony formation, cell death, expression of mitochondria-dependent pathway proteins, expression of EGFR, PI3K/AKT, MAPK signal pathway proteins, Bcl-2 mitochondrial translocation, ROS generation and cell apoptosis were examined by MTT, colony forming, live/dead cell staining, Western blot, immunofluorescence and flow cytometry assay.

Results: Our results showed that JB significantly induced cell growth inhibition and apoptotic cell death in PC-9, PC-9/GR and H1975 cells. JB activated mitochondria-mediated apoptotic pathway through inhibiting Bcl-2 mitochondrial translocation while inducing Bax translocated into mitochondria along with accumulated ROS production, thereby increasing the release of cytochrome c, subsequently cleaving procaspase9 into cleaved-caspase9 and then cleaving procaspase3 into cleaved-caspase3. Furthermore, the employment of protein kinase inhibitors LY294002 and SCH772984 revealed that the induction of mitochondria-mediated apoptosis by JB was reliant on inactivation of PI3K/AKT and MAPK signal pathways. Moreover, JB could synergize with gefitinib to induce apoptosis in PC-9, PC-9/GR and H1975 cells.

Conclusion: These data indicated that JB could be a potential therapeutic agent for NSCLC patients harboring EGFR mutations as well as those under gefitinib resistance.

Keywords: JuBei oral liquid; apoptosis; gefitinib; non-small cell lung cancer; resistance.

Grants and funding

The study was supported by grants from Postgraduate Innovative Project of Jiangsu Province (KYCX18_0817).