Wang Liguang, Wang Kai, Dong Wei, Shen Hongchang, Du Jiajun
Institute of Oncology, Shandong Provincial Hospital Affiliated to Shandong University, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, P.R. China.
Department of Oncology, Shandong Provincial Hospital Affiliated to Shandong University, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, P.R. China.
J Cancer. 2019 Jul 8;10(17):4085-4095. doi: 10.7150/jca.30012. eCollection 2019.
β-arrestin1 serves as scaffold proteins participating in multiple signaling pathways. However, there were few researches focusing on the impact of β-arrestin1 on DNA damage response (DDR). Non-small cell lung cancer cell (NSCLC) lines were transfected with β-arrestin1 plasmids or siRNA and received radiation treatment. MTT and colony formation assay were performed to assess the proliferation and viability of tumor cells. Flow cytometry was used to evaluate the impact of β-arrestin1 on radiation-induced apoptosis. Western blotting was applied to detect protein expression in apoptosis, DDR, ERK and NF-kB pathways. We used qRT-PCR to test ATR, H2AX, β-arrestin1 mRNA level in cancer tissues compared with para-carcinoma tissues. Co-IP was performed to evaluate the interaction between β-arrestin1 and ATR or H2AX. Comet assay was used to detect DNA damage. β-arrestin1 mRNA level co-related with ATR and H2AX levels in cancer tissues, and β-arrestin1 bound to ATR and H2AX directly or indirectly. Overexpression of β-arrestin1 enhanced the DNA damage response pathway activation and increase DNA damage and apoptosis. Interestingly, suppression of β-arrestin1 inhibited cell proliferation and attenuated ERK and NF-kB pathways activation induced by radiation. Overexpression of β-arrestin1 enhances DDR pathway activation induced by radiation, as well as downstream apoptosis, and depletion of β-arrestin1 inhibits DDR pathway. Meanwhile β-arrestin1 regulates cell proliferation by suppression of ERK and NF-kB pathways. Manipulation of β-arrestin1 status modulates radiosensitivity for NSCLC.
β-抑制蛋白1作为支架蛋白参与多种信号通路。然而，很少有研究关注β-抑制蛋白1对DNA损伤反应（DDR）的影响。用β-抑制蛋白1质粒或小干扰RNA转染非小细胞肺癌（NSCLC）细胞系，并进行放射治疗。采用MTT和集落形成试验评估肿瘤细胞的增殖和活力。用流式细胞术评估β-抑制蛋白1对辐射诱导凋亡的影响。采用蛋白质免疫印迹法检测凋亡、DDR、ERK和NF-κB信号通路中的蛋白表达。我们用qRT-PCR检测癌组织与癌旁组织中ATR、H2AX、β-抑制蛋白1的mRNA水平。采用免疫共沉淀法评估β-抑制蛋白1与ATR或H2AX之间的相互作用。用彗星试验检测DNA损伤。癌组织中β-抑制蛋白1的mRNA水平与ATR和H2AX水平呈正相关，且β-抑制蛋白1直接或间接与ATR和H2AX结合。β-抑制蛋白1的过表达增强了DNA损伤反应通路的激活，增加了DNA损伤和凋亡。有趣的是，抑制β-抑制蛋白1可抑制细胞增殖，并减弱辐射诱导的ERK和NF-κB信号通路的激活。β-抑制蛋白1的过表达增强了辐射诱导的DDR信号通路的激活以及下游凋亡，而β-抑制蛋白1的缺失则抑制DDR信号通路。同时，β-抑制蛋白1通过抑制ERK和NF-κB信号通路来调节细胞增殖。调控β-抑制蛋白1的状态可调节NSCLC的放射敏感性。
