Cao Fei, Ju Xiaoping, Chen Di, Jiang Lingong, Zhu Xiaofei, Qing Shuiwang, Fang Fang, Shen Yuxin, Jia Zhen, Zhang Huojun
Department of Radiation Oncology, Shanghai Changhai Hospital, Second Military Medical University, Shanghai 200433, P.R. China.
Mol Med Rep. 2017 Aug;16(2):2089-2094. doi: 10.3892/mmr.2017.6778. Epub 2017 Jun 14.
Emergence of resistance, unavoidable systemic toxicity and unsatisfactory efficacy arethe main obstacles for traditional cancer therapy. Combination with phosphorothioate modified antisense oligonucleotides (PS‑ASODN) against human telomerase reverse transcriptase (hTERT) may enhance the therapeutic effect of irradiation. However, the effect of PS‑ASODN against hTERT on the anti‑tumor effects of irradiation in liver cancer remain unclear. In the current study, Walker 256 cells were transfected with hTERT PS‑ASODN. Cell proliferation and cell viability were measured using the MTT assay and cell senescence was examined by SA‑β‑gal staining. Telomerase activity was determined by telomeric repeat amplification protocol‑polymerase chain reaction‑ELISA. Cell apoptosis was assayed by flow cytometry and DNA damage was determined by the comet assay.The PS‑ASODN was demonstrated to have an inhibitory effect on cell proliferation and accelerated effect on cell senescence by inhibiting telomerase activity. PS‑ASODN promoted the irradiation‑induced inhibition of cell viability and telomerase activity, and irradiation‑induced DNA damage and cell apoptosis via the activation of apoptosis‑associated proteins. Taken together, these results indicated that combined treatment of PS‑ASODN with irradiation significantly enhanced tumor inhibition. Therefore, PS‑ASODN provides an experimental foundation for gene therapy and is proposed for application in clinical treatment of liver cancer combined with radiotherapy.
耐药性的出现、不可避免的全身毒性和不尽人意的疗效是传统癌症治疗的主要障碍。与针对人类端粒酶逆转录酶（hTERT）的硫代磷酸酯修饰反义寡核苷酸（PS-ASODN）联合使用可能会增强放疗的治疗效果。然而，PS-ASODN针对hTERT对肝癌放疗抗肿瘤效果的影响仍不清楚。在本研究中，用hTERT PS-ASODN转染Walker 256细胞。使用MTT法测量细胞增殖和细胞活力，通过SA-β-半乳糖苷酶染色检测细胞衰老。通过端粒重复序列扩增协议-聚合酶链反应-酶联免疫吸附测定法测定端粒酶活性。通过流式细胞术检测细胞凋亡，通过彗星试验测定DNA损伤。PS-ASODN被证明通过抑制端粒酶活性对细胞增殖具有抑制作用，并对细胞衰老具有加速作用。PS-ASODN通过激活凋亡相关蛋白促进放疗诱导的细胞活力抑制和端粒酶活性抑制，以及放疗诱导的DNA损伤和细胞凋亡。综上所述，这些结果表明PS-ASODN与放疗联合治疗可显著增强肿瘤抑制作用。因此，PS-ASODN为基因治疗提供了实验基础，并被提议用于肝癌联合放疗的临床治疗。
