Wang Qiang, Wei Jianchang, Wang Chengxing, Zhang Tong, Huang Di, Wei Fang, He Feng, Cai Wensong, Yang Ping, Zeng Shanqi, Li Wanglin, Cao Jie
Department of General Surgery, Guangzhou Digestive Disease Centre, Guangzhou First People's Hospital, Guangzhou Medical University, Guangzhou, Guangdong 510180, P.R. China.
Department of Gastrointestinal Surgery, Jiangmen Central Hospital, Affiliated Jiangmen Hospital of Sun Yat-sen University, Jiangmen, Guangdong 529000, P.R. China.
Oncol Lett. 2018 Aug;16(2):2366-2372. doi: 10.3892/ol.2018.8916. Epub 2018 Jun 6.
Resistance to oxaliplatin (L-OHP) is a major obstacle to successful chemotherapy in colorectal cancer (CRC). In the present study, the ability of gambogic acid (GA) to reverse L-OHP resistance in CRC LoVo cells was investigated. L-OHP-resistant LoVo/L-OHP cells were established by exposing them to increasing concentrations of L-OHP. GA-reversed L-OHP-sensitive LoVo/L-OHP/GA cells were established by exposure to 0.5 µmol/l GA for 2 weeks. A Cell Counting Kit-8 assay was used to assess levels of proliferation. Flow cytometry was applied to detect apoptosis rates. Transwell assays were used to analyse invasion. Inductively coupled plasma mass spectrometry was used to determine intracellular platinum (Pt) content. Western blot analysis was used to reveal the protein levels of Human copper transporter 1 (hCTR1), Copper-transporting p-type adenosine triphosphatases 1 (ATP7A) and Copper-transporting p-type adenosine triphosphatases 2 (ATP7B). LoVo/L-OHP and LoVo/L-OHP/GA cell lines were successfully established, and it was identified that L-OHP inhibited the proliferation of LoVo, LoVo/L-OHP and LoVo/L-OHP/GA cells in a dose-dependent manner. Compared with the parent LoVo cells, the anti-apoptosis and invasion properties of LoVo/L-OHP cells were enhanced, and were reversed by GA treatment. Intracellular Pt content was highest in the LoVo cells, followed by LoVo/L-OHP/GA cells, and then lowest in the LoVo/L-OHP cells. Downregulated hCTP1 and upregulated ATP7A and ATP7B were associated with L-OHP resistance, and GA reversed the resistance by increasing levels of hCTR1 and decreasing levels of ATP7A and ATP7B. In conclusion, GA has the potential ability to reverse L-OHP resistance in CRC cells by increasing intracellular Pt content, which it achieves by increasing hCTR1 levels and decreasing ATP7A and ATP7B levels. GA may represent a promising treatment agent for L-OHP resistance.
对奥沙利铂(L-OHP)耐药是结直肠癌(CRC)化疗成功的主要障碍。在本研究中,研究了藤黄酸(GA)逆转CRC LoVo细胞中L-OHP耐药性的能力。通过将LoVo细胞暴露于浓度递增的L-OHP中建立L-OHP耐药的LoVo/L-OHP细胞。通过将细胞暴露于0.5 μmol/l GA中2周建立GA逆转的L-OHP敏感的LoVo/L-OHP/GA细胞。使用细胞计数试剂盒-8检测法评估增殖水平。应用流式细胞术检测凋亡率。使用Transwell检测法分析侵袭情况。使用电感耦合等离子体质谱法测定细胞内铂(Pt)含量。采用蛋白质印迹分析揭示人铜转运蛋白1(hCTR1)、铜转运P型ATP酶1(ATP7A)和铜转运P型ATP酶2(ATP7B)的蛋白水平。成功建立了LoVo/L-OHP和LoVo/L-OHP/GA细胞系,并且确定L-OHP以剂量依赖性方式抑制LoVo、LoVo/L-OHP和LoVo/L-OHP/GA细胞的增殖。与亲本LoVo细胞相比,LoVo/L-OHP细胞的抗凋亡和侵袭特性增强,而GA处理可使其逆转。细胞内Pt含量在LoVo细胞中最高,其次是LoVo/L-OHP/GA细胞,在LoVo/L-OHP细胞中最低。hCTP1下调以及ATP7A和ATP7B上调与L-OHP耐药相关,而GA通过增加hCTR1水平并降低ATP7A和ATP7B水平来逆转耐药性。总之,GA具有通过增加细胞内Pt含量来逆转CRC细胞中L-OHP耐药性的潜在能力,这是通过增加hCTR1水平并降低ATP7A和ATP7B水平实现的。GA可能是一种有前景的L-OHP耐药治疗药物。