Study on effect of DNA cross-link repair gene DCLRE1B on invasion and migration of hepatocellular carcinoma cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

DENG Zhaoda ^1,2 , WANG Jing ² , LI Lincheng ^1,2 , ZOU Wenbo ^1,2 ,  LIU Rong ^1,2

1. Faculty of Hepato-Biliary-Pancreatic Surgery, Chinese People’s Liberation Army General Hospital, Beijing 100853, P. R. China;
2. Chinese People’s Liberation Army Medicine School, Beijing 100853, P. R. China;

Corresponding?author：

LIU Rong, Email: liurong301@126.com

Keywords：

DNA cross-linking repair; hepatocellular carcinoma; invasion; migration; biological information

DOI：

10.7507/1007-9424.202111066

Video：

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Objective To explore the effects of DNA cross-linking repair 1B (DCLRE1B) gene on the migration and invasion ability of hepatocellular carcinoma cell. Methods Bioinformatics analysis was used to analyze the expression of DCLRE1B mRNA in hepatocellular carcinoma, and its relationship with the prognosis and related influencing factors of patients. Immunohistochemical staining was used to detect the expression of DCLRE1B protein in resected hepatocellular carcinoma tissues and their corresponding normal liver tissues. The DCLRE1B gene silenced Huh7 and HepG2 hepatocellular carcinoma cell lines were constructed by lentivirus, and the transfected effect was detected by Western blot. The migration and invasion of DCLRE1B silenced hepatocellular carcinoma cells were detected by scratch test and Transwell method. The changes of genes related to epithelial mesenchymal transformation (EMT) after DCLRE1B silencing were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Results ① The biological information analysis results showed that: The mRNA expression of DCLRE1B was highly expressed in a variety of tumors including hepatocellular carcinoma (P<0.05). The mRNA expression of DCLRE1B was associated with the TNM staging of tumor (P<0.05). The relative expression level of DCLRE1B mRNA in hepatocellular carcinoma patients was related to their prognosis. The overall survival situation (P=0.038) and progression free survival situation (P=0.005) of hepatocellular carcinoma patients in the high expression group were worse than those in the low expression group. Univariate and multivariate Cox analysis showed that the expression of DCLRE1B gene was an independent factor affecting the prognosis of hepatocellular carcinoma (P<0.05). ② The positive rate of DCLRE1B protein expression in resected hepatocellular carcinoma tissues was higher than that in normal liver tissues (P<0.05). ③ Cell experiment results showed that: After stable silencing DCLRE1B gene of hepatocellular carcinoma cell (Huh7 and HepG2) constructed by lentivirus, the expression of DCLRE1B protein was significantly down regulated (P<0.05). After silencing DCLRE1B gene, the migration and invasion ability of hepatocellular carcinoma cells were significantly decreased (P<0.05). After silencing DCLRE1B, the mRNA expressions of E-cadherin, matrix metalloproteinase 9, and β-catenin were up regulated (P<0.05), and the mRNA expressions of N-cadherin and Vimentin were down regulated (P<0.05), but the mRNA expression of zinc finger transcription factor had no significant change, and the difference was not statistically significant (P＞0.05). Conclusion Silencing DCLRE1B gene can inhibit the migration and invasion ability of hepatocellular carcinoma cells, and its mechanism may be related to the process of EMT.

Citation： DENG Zhaoda, WANG Jing, LI Lincheng, ZOU Wenbo, LIU Rong. Study on effect of DNA cross-link repair gene DCLRE1B on invasion and migration of hepatocellular carcinoma cells. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2022, 29(5): 585-593. doi: 10.7507/1007-9424.202111066 Copy

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2.	Rycenga HB, Long DT. The evolving role of DNA inter-strand crosslinks in chemotherapy. Curr Opin Pharmacol, 2018, 41: 20-26.
3.	Basourakos SP, Li L, Aparicio AM, et al. Combination platinum-based and DNA damage response-targeting cancer therapy: evolution and future directions. Curr Med Chem, 2017, 24(15): 1586-1606.
4.	Nikolova T, Roos WP, Kr?mer OH, et al. Chloroethylating nitrosoureas in cancer therapy: DNA damage, repair and cell death signaling. Biochim Biophys Acta Rev Cancer, 2017, 1868(1): 29-39.
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10.	Wang H, Xiang D, Liu B, et al. Inadequate DNA damage repair promotes mammary transdifferentiation, leading to BRCA1 breast cancer. Cell, 2019, 178(1): 135-151.
11.	Guohui S, Lijiao Z, Rugang Z. The induction and repair of DNA interstrand crosslinks and implications in cancer chemotherapy. Anticancer Agents Med Chem, 2015, 16(2): 221-246.
12.	Bae JB, Mukhopadhyay SS, Liu L, et al. Snm1B/Apollo mediates replication fork collapse and S phase checkpoint activation in response to DNA interstrand cross-links. Oncogene, 2008, 27(37): 5045-5056.
13.	Ishiai M, Kimura M, Namikoshi K, et al. DNA cross-link repair protein SNM1A interacts with PIAS1 in nuclear focus formation. Mol Cell Biol, 2004, 24(24): 10733-10741.
14.	Demuth I, Digweed M, Concannon P. Human SNM1B is required for normal cellular response to both DNA interstrand crosslink-inducing agents and ionizing radiation. Oncogene, 2004, 23(53): 8611-8618.
15.	Mason JM, Sekiguchi JM. Snm1B/Apollo functions in the Fanconi anemia pathway in response to DNA interstrand crosslinks. Hum Mol Genet, 2011, 20(13): 2549-2559.
16.	Jin MH, Oh DY. ATM in DNA repair in cancer. Pharmacol Ther, 2019, 203: 107391. doi: 10.1016/j.pharmthera.2019.07.002.
17.	Mason JM, Das I, Arlt M, et al. The SNM1B/Apollo DNA nuclease functions in resolution of replication stress and maintenance of common fragile site stability. Hum Mol Genet, 2013, 22(24): 4901-4913.
18.	Karami S, Han Y, Pande M, et al. Telomere structure and maintenance gene variants and risk of five cancer types. Int J Cancer, 2016, 139(12): 2655-2670.
19.	Lee YK, Lim J, Yoon SY, et al. Promotion of cell death in cisplatin-resistant ovarian cancer cells through KDM1B-DCLRE1B modulation. Int J Mol Sci, 2019, 20(10): 2443. doi: 10.3390/ijms20102443.
20.	Baddock HT, Newman JA, Yosaatmadja Y, et al. A phosphate binding pocket is a key determinant of exo- versus endo-nucleolytic activity in the SNM1 nuclease family. Nucleic Acids Res, 2021, 49(16): 9294-9309.
21.	Yosaatmadja Y, Baddock HT, Newman JA, et al. Structural and mechanistic insights into the Artemis endonuclease and strategies for its inhibition. Nucleic Acids Res, 2021, 49(16): 9310-9326.
22.	Lobo-Alves SC, de Oliveira LA, Petzl-Erler ML. Region 1p132 including the RSBN1, PTPN22, AP4B1 and long non-coding RNA genes does not bear risk factors for endemic pemphigus foliaceus (fogo selvagem). Int J Immunogenet, 2019, 46(3): 139-145.
23.	Stringer JR, Counter CM. Snm1B interacts with PSF2. PLoS One, 2012, 7(11): e49626. doi: 10.1371/journal.pone.0049626.
24.	Jiang X, Wang L, Xie S, et al. Long noncoding RNA MEG3 blocks telomerase activity in human liver cancer stem cells epigenetically. Stem Cell Res Ther, 2020, 11(1): 518. doi: 10.1186/s13287-020-02036-4.
25.	Montanari M, Rossetti S, Cavaliere C, et al. Epithelial-mesenchymal transition in prostate cancer: an overview. Oncotarget, 2017, 8(21): 35376-35389.
26.	Lu W, Kang Y. Epithelial-mesenchymal plasticity in cancer progression and metastasis. Dev Cell, 2019, 49(3): 361-374.
27.	Kim YN, Koo KH, Sung JY, et al. Anoikis resistance: an essential prerequisite for tumor metastasis. Int J Cell Biol, 2012, 2012: 306879. doi: 10.1155/2012/306879 .
28.	Aiello NM, Kang Y. Context-dependent EMT programs in cancer metastasis. J Exp Med, 2019, 216(5): 1016-1026.

1. Baddock HT, Yosaatmadja Y, Newman JA, et al. The SNM1A DNA repair nuclease. DNA Repair (Amst), 2020, 95: 102941. doi: 10.1016/j.dnarep.2020.102941.
2. Rycenga HB, Long DT. The evolving role of DNA inter-strand crosslinks in chemotherapy. Curr Opin Pharmacol, 2018, 41: 20-26.
3. Basourakos SP, Li L, Aparicio AM, et al. Combination platinum-based and DNA damage response-targeting cancer therapy: evolution and future directions. Curr Med Chem, 2017, 24(15): 1586-1606.
4. Nikolova T, Roos WP, Kr?mer OH, et al. Chloroethylating nitrosoureas in cancer therapy: DNA damage, repair and cell death signaling. Biochim Biophys Acta Rev Cancer, 2017, 1868(1): 29-39.
5. Munari FM, Guecheva TN, Bonatto D, et al. New features on Pso2 protein family in DNA interstrand cross-link repair and in the maintenance of genomic integrity in saccharomyces cerevisiae. Fungal Genet Biol, 2013, 60: 122-132.
6. Demuth I, Bradshaw PS, Lindner A, et al. Endogenous hSNM1B/Apollo interacts with TRF2 and stimulates ATM in response to ionizing radiation. DNA Repair (Amst), 2008, 7(8): 1192-1201.
7. van Overbeek M, de Lange T. Apollo, an Artemis-related nuclease, interacts with TRF2 and protects human telomeres in S phase. Curr Biol, 2006, 16(13): 1295-1302.
8. Giannelli G, Koudelkova P, Dituri F, et al. Role of epithelial to mesenchymal transition in hepatocellular carcinoma. J Hepatol, 2016, 65(4): 798-808.
9. Enderle J, Dorn A, Puchta H. DNA- and DNA-protein-crosslink repair in plants. Int J Mol Sci, 2019, 20(17): 4304. doi: 10.3390/ijms20174304.
10. Wang H, Xiang D, Liu B, et al. Inadequate DNA damage repair promotes mammary transdifferentiation, leading to BRCA1 breast cancer. Cell, 2019, 178(1): 135-151.
11. Guohui S, Lijiao Z, Rugang Z. The induction and repair of DNA interstrand crosslinks and implications in cancer chemotherapy. Anticancer Agents Med Chem, 2015, 16(2): 221-246.
12. Bae JB, Mukhopadhyay SS, Liu L, et al. Snm1B/Apollo mediates replication fork collapse and S phase checkpoint activation in response to DNA interstrand cross-links. Oncogene, 2008, 27(37): 5045-5056.
13. Ishiai M, Kimura M, Namikoshi K, et al. DNA cross-link repair protein SNM1A interacts with PIAS1 in nuclear focus formation. Mol Cell Biol, 2004, 24(24): 10733-10741.
14. Demuth I, Digweed M, Concannon P. Human SNM1B is required for normal cellular response to both DNA interstrand crosslink-inducing agents and ionizing radiation. Oncogene, 2004, 23(53): 8611-8618.
15. Mason JM, Sekiguchi JM. Snm1B/Apollo functions in the Fanconi anemia pathway in response to DNA interstrand crosslinks. Hum Mol Genet, 2011, 20(13): 2549-2559.
16. Jin MH, Oh DY. ATM in DNA repair in cancer. Pharmacol Ther, 2019, 203: 107391. doi: 10.1016/j.pharmthera.2019.07.002.
17. Mason JM, Das I, Arlt M, et al. The SNM1B/Apollo DNA nuclease functions in resolution of replication stress and maintenance of common fragile site stability. Hum Mol Genet, 2013, 22(24): 4901-4913.
18. Karami S, Han Y, Pande M, et al. Telomere structure and maintenance gene variants and risk of five cancer types. Int J Cancer, 2016, 139(12): 2655-2670.
19. Lee YK, Lim J, Yoon SY, et al. Promotion of cell death in cisplatin-resistant ovarian cancer cells through KDM1B-DCLRE1B modulation. Int J Mol Sci, 2019, 20(10): 2443. doi: 10.3390/ijms20102443.
20. Baddock HT, Newman JA, Yosaatmadja Y, et al. A phosphate binding pocket is a key determinant of exo- versus endo-nucleolytic activity in the SNM1 nuclease family. Nucleic Acids Res, 2021, 49(16): 9294-9309.
21. Yosaatmadja Y, Baddock HT, Newman JA, et al. Structural and mechanistic insights into the Artemis endonuclease and strategies for its inhibition. Nucleic Acids Res, 2021, 49(16): 9310-9326.
22. Lobo-Alves SC, de Oliveira LA, Petzl-Erler ML. Region 1p132 including the RSBN1, PTPN22, AP4B1 and long non-coding RNA genes does not bear risk factors for endemic pemphigus foliaceus (fogo selvagem). Int J Immunogenet, 2019, 46(3): 139-145.
23. Stringer JR, Counter CM. Snm1B interacts with PSF2. PLoS One, 2012, 7(11): e49626. doi: 10.1371/journal.pone.0049626.
24. Jiang X, Wang L, Xie S, et al. Long noncoding RNA MEG3 blocks telomerase activity in human liver cancer stem cells epigenetically. Stem Cell Res Ther, 2020, 11(1): 518. doi: 10.1186/s13287-020-02036-4.
25. Montanari M, Rossetti S, Cavaliere C, et al. Epithelial-mesenchymal transition in prostate cancer: an overview. Oncotarget, 2017, 8(21): 35376-35389.
26. Lu W, Kang Y. Epithelial-mesenchymal plasticity in cancer progression and metastasis. Dev Cell, 2019, 49(3): 361-374.
27. Kim YN, Koo KH, Sung JY, et al. Anoikis resistance: an essential prerequisite for tumor metastasis. Int J Cell Biol, 2012, 2012: 306879. doi: 10.1155/2012/306879 .
28. Aiello NM, Kang Y. Context-dependent EMT programs in cancer metastasis. J Exp Med, 2019, 216(5): 1016-1026.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Study on effect of DNA cross-link repair gene DCLRE1B on invasion and migration of hepatocellular carcinoma cells

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