Sakhawat Ali, Liu Yanan, Ma Ling, Muhammad Tahir, Wang Shensen, Zhang Lina, Cong Xianling, Huang Yinghui
College of Life Sciences and Bio-Engineering, Beijing University of Technology, China.
Basic Medical College, Jilin University, China.
J Cancer. 2017 May 12;8(8):1425-1432. doi: 10.7150/jca.18371. eCollection 2017.
Conditionally replicating adenoviruses (CRAds) have been proven potent oncolytic viruses in previous studies. They selectively replicate in the tumor cells because of incorporated survivin promoter and ultimately lead to their killing with minimal side effects on normal tissue. Chemotherapy with cisplatin is commonly employed for treating tumors, but its cytotoxic effects and development of resistance remained major concerns to be dealt with. The aim of this study was to explore the anticancer potential of survivin regulated CRAd alone or in combination with cisplatin in the A549 lung cancer cell line and cisplatin-resistant lung cancer cell line, A549-DDPR. CRAd was genetically engineered in our laboratory by removing its E1B region and adding survivin promoter to control its replication. A549, H292, and H661 lung cancer cell lines were procured from the CAS-China. The anti-tumor effectiveness of combined treatment (cisplatin plus CRAd) was evaluated in vitro through MTS assays and in vivo through mouse model experimentation. RT- PCR was used to assess MDR gene and mRNA expression of coxsackie adenoviral receptor (CAR). Results of studies established that A549 lung cancer cells were highly sensitive to cisplatin showing dose-dependent inhibition. The resistant cells of A549-DDPR exhibited very less sensitivity to cisplatin but were infected with CRAd more efficiently as compared to A549. A549-DDPR cells exhibited higher expression of MDR gene and CAR in the RT-PCR analysis. The nearly similar rise in the CAR expression was seen when lung cancer cell lines received cisplatin in combined treatment (cisplatin plus CRAd). Combined anti-cancer therapy (cisplatin plus oncolytic virus) proved more efficient than monotherapy in the killing of cancer cells. Results of experiments recapitulated nearly similar tumor inhibition activities. This study highlighted the significant role of survivin in gene therapy as it has the potential to render CRAd more tumor specific. It also establishes that higher CAR expression plays a vital role in the success of adenovirus-based therapies. Furthermore, a careful combination of chemotherapy drugs and oncolytic viruses can culminate in significant therapeutic achievements against cancer.
在先前的研究中，条件性复制腺病毒（CRAds）已被证明是有效的溶瘤病毒。由于整合了生存素启动子，它们在肿瘤细胞中选择性复制，并最终导致肿瘤细胞死亡，同时对正常组织的副作用最小。顺铂化疗常用于治疗肿瘤，但其细胞毒性作用和耐药性的产生仍是需要解决的主要问题。本研究的目的是探讨生存素调控的CRAd单独或与顺铂联合在A549肺癌细胞系和顺铂耐药肺癌细胞系A549-DDPR中的抗癌潜力。CRAd是我们实验室通过去除其E1B区域并添加生存素启动子来控制其复制而进行基因工程改造的。A549、H292和H661肺癌细胞系购自中国科学院。联合治疗（顺铂加CRAd）的抗肿瘤效果通过MTS试验在体外进行评估，并通过小鼠模型实验在体内进行评估。RT-PCR用于评估多药耐药基因和柯萨奇腺病毒受体（CAR）的mRNA表达。研究结果表明，A549肺癌细胞对顺铂高度敏感，呈剂量依赖性抑制。A549-DDPR的耐药细胞对顺铂的敏感性很低，但与A549相比，它们被CRAd感染的效率更高。在RT-PCR分析中，A549-DDPR细胞的多药耐药基因和CAR表达较高。当肺癌细胞系接受联合治疗（顺铂加CRAd）时，CAR表达出现了几乎相似的升高。联合抗癌治疗（顺铂加溶瘤病毒）在杀死癌细胞方面比单一疗法更有效。实验结果概括出了几乎相似的肿瘤抑制活性。这项研究突出了生存素在基因治疗中的重要作用，因为它有可能使CRAd更具肿瘤特异性。它还表明，较高的CAR表达在基于腺病毒的治疗成功中起着至关重要的作用。此外，化疗药物和溶瘤病毒的精心组合可以在抗癌治疗中取得显著的治疗成果。
