Autophagy-related Proteins as a Prognostic Factor of Patients With Colorectal Cancer
Autophagy-related Proteins as a Prognostic Factor of Patients With Colorectal Cancer
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DOI：
10.1097/coc.0000000000000592
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发表时间：
2019-10-01
影响因子：
2.6
通讯作者：
Karamouzis, Michalis V.
中科院分区：
文献类型：
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作者：
Koustas, Evangelos;Sarantis, Panagiotis;Karamouzis, Michalis V.
目的：自噬在肿瘤发生中起双重作用。在初始阶段，它促进细胞存活并抑制癌发生，而在癌症发展中，它诱导癌细胞存活。在这项研究中，我们研究了自噬在结直肠癌（CRC）细胞系中作为保护或肿瘤抑制机制的作用，并评估了其作为人类肿瘤样本中潜在生物标志物的作用。材料与方法：对2016年1月1日至12月31日在我科治疗的68例结直肠癌患者的资料进行分析。对福尔马林固定的石蜡包埋组织样本中的p62、LC 3B、Beclin-1和Rab-7进行了免疫组织化学评估，其表达与临床病理特征、突变状态和治疗方法相关。卡方检验用于检验分类变量之间的关联。使用Kaplan-Meier方法估计生存曲线，并使用对数秩检验评估差异。还使用了科洛-205、HT 29、SW-480和Caco-2细胞系，以检测奥沙利铂、伊立替康、羟氯喹和3-甲基腺嘌呤的自噬标志物。结果如下：Beclin-1的过表达与接受化疗的CRC患者的生存率差相关（P=0.001），与分期和突变状态无关。Rab-7也与无进展生存期（PFS）相关（P=0.088）。奥沙利铂（10和20 μ M）和伊立替康（10和20 μ M）抑制微卫星稳定（MSS）CRC细胞系中的自噬。通过单丹酰尸胺染色进一步鉴定用奥沙利铂和伊立替康处理后MSS CRC细胞系中自噬的抑制。此外，通过p62和LC 3B的积累鉴定了用诸如羟氯喹（20 μ M）和3-甲基腺嘌呤（5 mM）的分子抑制自噬。结论：Beclin-1是影响总生存期和PFS的独立预后因素。此外，Rab-7被确定为PFS的独立预后因素。此外，几种化疗药物如奥沙利铂和伊立替康以类似于羟氯喹和3-甲基腺嘌呤的方式抑制MSS CRC细胞系中的自噬。因此，在出现化疗耐药性的MSS患者中，包括自噬抑制剂在内的其他疗法的组合可能更有益。需要进一步的临床试验来研究这些治疗策略。
Objectives: Autophagy plays a dual role in tumorigenesis. In the initial stages, it promotes cell survival and suppresses carcinogenesis, whereas in cancer development, it induces cancer cell survival. In this study, we investigate the role of autophagy as a protective or tumor suppressor mechanism in colorectal cancer (CRC) cell lines and evaluate its role as a potential biomarker in human tumor samples. Materials and Methods: The data of 68 patients with CRC treated at our Department from January 1 to December 31, 2016 were analyzed. Immunohistochemistry evaluation of p62, LC3B, Beclin-1, and Rab-7 in formalin-fixed paraffin-embedded tissue samples was performed and their expression was correlated with clinicopathologic characteristics, mutation status, and therapeutic approach. The chi(2) was used to test an association among categorical variables. Survival curves were estimated using the Kaplan-Meier method and differences were assessed using the log-rank test. Colo-205, HT29, SW-480, and Caco-2 cell lines were also used so as to test the autophagy markers with oxaliplatin, irinotecan, hydroxychloroquine, and 3-methyladenine. Results: Overexpression of Beclin-1 is associated with poor survival (P=0.001) in patients with CRC treated with chemotherapy, irrespective of the stage and mutational status. Rab-7 is also correlated with progression-free survival (PFS) (P=0.088). Oxaliplatin (10 and 20 mu M) and irinotecan (10 and 20 mu M) inhibit autophagy in microsatellite stable (MSS) CRC cell lines. The inhibition of autophagy in MSS CRC cell lines after treatment with oxaliplatin and irinotecan is further identified through monodancylcadaverine staining. Moreover, inhibition of autophagy with molecules such as hydroxychloroquine (20 mu M) and 3-methyladenine (5 mM) was identified by the accumulation of p62 and LC3B. Conclusions: Beclin-1 is an independent prognostic factor of overall survival and PFS. Also, Rab-7 is identified as an independent prognostic factor of PFS. Besides, several chemotherapeutic drugs such as oxaliplatin and irinotecan inhibit autophagy in MSS CRC cell lines in a similar way like hydroxychloroquine and 3-methyladenine. Thus, in MSS patients who develop chemoresistance, a combination of other therapies that include an autophagy inhibitor could be more beneficial. Further clinical trials are needed to investigate these therapeutic strategies.