DNASE1L3 inhibits hepatocellular carcinoma by delaying cell cycle progression through CDK2.
DNASE1L3 inhibits hepatocellular carcinoma by delaying cell cycle progression through CDK2.
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DOI：
10.1007/s13402-022-00709-1
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发表时间：
2022-11
影响因子：
6.6
通讯作者：
Jiaqi Sun;Xiyang Wang;Qingsong Shen;Min Wang;Shuxian Chen;Xuechun Zhang;Yongping Huang;Zhongling Zhang;Wenhua Li;Yufeng Yuan;Zan Huang
中科院分区：
文献类型：
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作者：
Jiaqi Sun;Xiyang Wang;Qingsong Shen;Min Wang;Shuxian Chen;Xuechun Zhang;Yongping Huang;Zhongling Zhang;Wenhua Li;Yufeng Yuan;Zan Huang
目的：细胞周期靶向失调是治疗肝细胞癌（HCC）的一种行之有效的治疗策略。剖析其潜在机制可能提高HCC治疗的疗效。方法利用TCGA和新临床样本的shcc数据进行DNASE1L3表达分析，并评估其与HCC发展的相关性。采用RTCA、CCK-8和transwell法检测DNASE1L3在体外对HCC细胞增殖、集落形成、迁移和侵袭的作用，以及在体内对异种移植裸鼠模型皮下肿瘤形成的作用。在AKT/ nrasv12诱导和DEN/ ccl4诱导的原发性肝癌野生型和DNASE1L3−/−小鼠中，进一步验证了DNASE1L3在HCC肿瘤发生中的作用。最后，通过RNA-Seq分析以及细胞周期、免疫荧光、共免疫沉淀和Western blotting等生化方法揭示其潜在机制。结果我们发现DNASE1L3在HCC中显著下调，是一个有利的预后因素。DNASE1L3在体外显著减弱HCC细胞增殖、集落形成、迁移和侵袭，在体内显著减少裸鼠皮下肿瘤形成。此外，DNASE1L3过表达抑制了AKT/ nrasv12诱导的野生型小鼠肝癌，DNASE1L3缺乏加重了DEN/ ccl4诱导的小鼠肝癌。系统分析显示，DNASE1L3通过与CDK2相互作用并抑制CDK2刺激的E2F1活性来损害HCC细胞周期进展。c端缺失（DNASE1L3ΔCT）减少了与CDK2的相互作用，并取消了对HCC的抑制功能。结论我们的研究揭示了DNASE1L3是一种新的肝癌细胞周期调节因子和肿瘤抑制因子。DNASE1L3可能通过破坏阳性的E2F1-CDK2调控环而延缓细胞周期进程，从而损害HCC的肿瘤发生。DNASE1L3可能作为肝癌新治疗策略开发的靶点。
PurposeDysregulated cell cycle targeting is a well-established therapeutic strategy against hepatocellular carcinoma (HCC). Dissecting the underlying mechanism may improve the efficacy of HCC therapy.MethodsHCC data from TCGA and new clinical samples were used for DNASE1L3 expression analysis and for assessing its correlation with HCC development. The in vitro function of DNASE1L3 in HCC cell proliferation, colony formation, migration and invasion was assessed using RTCA, CCK-8 and transwell assays and the in vivo function in subcutaneous tumor formation in a xenograft nude mouse model. The role of DNASE1L3 in HCC tumorigenesis was further verified in AKT/NRASV12-induced and DEN/CCl4-induced primary liver cancers in wildtype andDnase1l3−/−mice. Finally, RNA-Seq analysis followed by biochemical methods including cell cycle, immunofluorescence, co-immunoprecipitation and Western blotting assays were employed to reveal the underlying mechanism.ResultsWe found that DNASE1L3 was significantly downregulated and served as a favorable prognostic factor in HCC. DNASE1L3 dramatically attenuated HCC cell proliferation, colony formation, migration and invasion in vitro and reduced subcutaneous tumor formation in nude mice in vivo. Furthermore, DNASE1L3 overexpression dampened AKT/NRASV12-induced mouse liver cancer in wildtype mice and DNASE1L3 deficiency worsened DEN/CCl4-induced liver cancer inDnase1l3−/−mice. Systemic analysis revealed that DNASE1L3 impaired HCC cell cycle progression by interacting with CDK2 and inhibiting CDK2-stimulated E2F1 activity. C-terminal deletion (DNASE1L3ΔCT) diminished the interaction with CDK2 and abrogated the inhibitory function against HCC.ConclusionOur study unveils DNASE1L3 as a novel HCC cell cycle regulator and tumor suppressor. DNASE1L3 impairs HCC tumorigenesis by delaying cell cycle progression possibly through disrupting the positive E2F1-CDK2 regulatory loop. DNASE1L3 may serve as a target for the development of novel therapeutic strategies against HCC.