Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Transcription Factor E2F7 Hampers the Killing Effect of NK Cells against Colorectal Cancer Cells via Activating RAD18 Transcription

  • Bingdong Jiang (Department of Oncology, Union Jiangbei Hospital Huazhong University of Science and Technology) ;
  • Binghua Yan (Department of Radiation Oncology, Huai'an Hospital of Huai'an City) ;
  • Hengjin Yang (Department of Radiation Oncology, Huai'an Hospital of Huai'an City) ;
  • He Geng (Department of Radiation Oncology, Huai'an Hospital of Huai'an City) ;
  • Peng Li (Department of Radiation Oncology, Huai'an Hospital of Huai'an City)
  • Received : 2023.08.16
  • Accepted : 2023.11.07
  • Published : 2024.04.28
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Abstract

As a pivotal defensive line against multitudinous malignant tumors, natural killer (NK) cells exist in the tumor microenvironment (TME). RAD18 E3 Ubiquitin Protein Ligase (RAD18) has been reported to foster the malignant progression of multiple cancers, but its effect on NK function has not been mined. Here, the study was designed to mine the mechanism by which RAD18 regulates the killing effect of NK cells on colorectal cancer (CRC) cells. Expression of E2F Transcription Factor 7 (E2F7) and RAD18 in CRC tissues, their correlation, binding sites, and RAD18 enrichment pathway were analyzed by bioinformatics. Expression of E2F7 and RAD18 in cells was assayed by qRT-PCR and western blot. Dual-luciferase assay and chromatin immunoprecipitation (ChIP) assay verified the regulatory relationship between E2F7 and RAD18. CCK-8 assay was utilized to assay cell viability, colony formation assay to detect cell proliferation, lactate dehydrogenase (LDH) test to assay NK cell cytotoxicity, ELISA to assay levels of granulocyte-macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ), and immunofluorescence to detect expression of toxic molecules perforin and granzyme B. High expression of RAD18 and E2F7 was found in CRC tissues and cells. Silencing RAD18 could hamper the proliferation of CRC cells, foster viability and cytotoxicity of NK cells, and increase the secretion of GM-CSF, TNF-α, IFN-γ as well as the expression of perforin and granzyme B. Additionally, ChIP and dual-luciferase reporter assay ascertained the binding relationship between RAD18 promoter region and E2F7. E2F7 could activate the transcription of RAD18, and silencing RAD18 reversed the inhibitory effect of E2F7 overexpression on NK cell killing. This work clarified the inhibitory effect of the E2F7/RAD18 axis on NK cell killing in CRC, and proffered a new direction for immunotherapy of CRC in targeted immune microenvironment.

Keywords

Acknowledgement

This study was supported by the funds from Huai'an Science and Technology Project (HAB202138). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

References

  1. Siegel RL, Miller KD, Fuchs HE, Jemal A, Cancer statistics, 2021. CA Cancer J. Clin. 71: 7-33. 
  2. Siegel RL, Miller KD, Goding Sauer A, Fedewa SA, Butterly LF, Anderson JC, et al. 2020. Colorectal cancer statistics, 2020. CA Cancer J. Clin. 70: 145-164. 
  3. Johdi NA, Sukor NF. 2020. Colorectal cancer immunotherapy: options and strategies. Front. Immunol. 11: 1624. 
  4. Fan A, Wang B, Wang X, Nie Y, Fan D, Zhao X, et al. 2021. Immunotherapy in colorectal cancer: current achievements and future perspective. Int. J. Biol. Sci. 17: 3837-3849. 
  5. Gomez-Lomeli P, Bravo-Cuellar A, Hernandez-Flores G, Jave-Suarez LF, Aguilar-Lemarroy A, Lerma-Diaz JM, et al. 2014. Increase of IFN-gamma and TNF-alpha production in CD107a+ NK-92 cells co-cultured with cervical cancer cell lines pre-treated with the HO-1 inhibitor. Cancer Cell Int. 14: 100. 
  6. Zhang Z, Liu L, Cao S, Zhu Y, Mei Q. 2017. Gene delivery of TIPE2 inhibits breast cancer development and metastasis via CD8(+) T and NK cell-mediated antitumor responses. Mol. Immunol. 85: 230-237. 
  7. Huang Y, Luo Y, Ou W, Wang Y, Dong D, Peng X., et al. 2021. Exosomal lncRNA SNHG10 derived from colorectal cancer cells suppresses natural killer cell cytotoxicity by upregulating INHBC. Cancer Cell Int. 21: 528. 
  8. Tang S, Fu H, Xu Q, Zhou Y. 2019. miR-20a regulates sensitivity of colorectal cancer cells to NK cells by targeting MICA. Biosci. Rep. 39: BSR20180695. 
  9. Huang J, Huen MS, Kim H, Leung CC, Glover JN, Yu X, et al. 2009. RAD18 transmits DNA damage signalling to elicit homologous recombination repair. Nat. Cell Biol. 11: 592-603. 
  10. Lou P, Zou S, Shang Z, He C, Gao A, Hou S, et al. 2019. RAD18 contributes to the migration and invasion of human cervical cancer cells via the interleukin-1beta pathway. Mol. Med. Rep. 20: 3415-3423. 
  11. Zou S, Yang J, Guo J, Su Y, He C, Wu J, et al. 2018. RAD18 promotes the migration and invasion of esophageal squamous cell cancer via the JNK-MMPs pathway. Cancer Lett. 417: 65-74. 
  12. Xue H, Wu Z, Rao D, Zhuo B, Chen Q. 2020. Long non-coding RNA LINC00858 aggravates the oncogenic phenotypes of ovarian cancer cells through miR-134-5p/RAD18 signaling. Arch. Gynecol. Obstet. 302: 1243-1254. 
  13. Baatar S, Bai T, Yokobori T, Gombodorj N, Nakazawa N, Ubukata Y, et al. 2020. High RAD18 expression is associated with disease progression and poor prognosis in patients with gastric cancer. Ann. Surg. Oncol. 27: 4360-4368. 
  14. Li YP, Du XR, Zhang R, Yang Q. 2021. Interleukin-18 promotes the antitumor ability of natural killer cells in colorectal cancer via the miR-574-3p/TGF-beta1 axis. Bioengineered 12: 763-778. 
  15. Fang P, Xiang L, Chen W, Li S, Huang S, Li J, et al. 2019. LncRNA GAS5 enhanced the killing effect of NK cell on liver cancer through regulating miR-544/RUNX3. Innate Immun. 25: 99-109. 
  16. Sato N, Sakai N, Furukawa K, Takayashiki T, Kuboki S, Takano S, et al. 2022. Tumor-suppressive role of Smad ubiquitination regulatory factor 2 in patients with colorectal cancer. Sci. Rep. 12: 5495. 
  17. Tang J, Yang L, Li Y, Ning X, Chaulagain A, Wang T, et al. 2021. ARID3A promotes the development of colorectal cancer by upregulating AURKA. Carcinogenesis 42: 578-586. 
  18. Konstantinidis KV, Alici E, Aints A, Christensson B, Ljunggren HG, Dilber M. S. 2005. Targeting IL-2 to the endoplasmic reticulum confines autocrine growth stimulation to NK-92 cells. Exp. Hematol. 33: 159-164. 
  19. Wong SH, Yu J. 2019. Gut microbiota in colorectal cancer: mechanisms of action and clinical applications. Nat. Rev. Gastroenterol. Hepatol. 16: 690-704. 
  20. Allen J, Sears CL. 2019. Impact of the gut microbiome on the genome and epigenome of colon epithelial cells: contributions to colorectal cancer development. Genome Med. 11: 11. 
  21. Yan X, Chen J, Meng Y, He C, Zou S, Li P, et al. 2019. RAD18 may function as a predictor of response to preoperative concurrent chemoradiotherapy in patients with locally advanced rectal cancer through caspase-9-caspase-3-dependent apoptotic pathway. Cancer Med. 8: 3094-3104. 
  22. Lou P, Sun X, Zhou J, Zou S. 2016. [Effect of RAD18-siRNA on proliferation and chemotherapy sensitivity of human esophageal squamous cell carcinoma ECA-109 cells]. Zhejiang Da Xue Xue Bao Yi Xue Ban 45: 364-370. 
  23. Liu RL, Dong Y, Deng YZ, Wang WJ, Li WD. 2015. Tumor suppressor miR-145 reverses drug resistance by directly targeting DNA damage-related gene RAD18 in colorectal cancer. Tumour Biol. 36: 5011-5019. 
  24. Li P, He C, Gao A, Yan X, Xia X, Zhou J, et al. 2020. RAD18 promotes colorectal cancer metastasis by activating the epithelial-mesenchymal transition pathway. Oncol. Rep. 44: 213-223. 
  25. Bhat R, Rommelaere J. 2013. NK-cell-dependent killing of colon carcinoma cells is mediated by natural cytotoxicity receptors (NCRs) and stimulated by parvovirus infection of target cells. BMC Cancer 13: 367. 
  26. Abdelrahman MM, Fawzy IO, Bassiouni AA, Gomaa AI, Esmat G, Waked I, et al. 2016. Enhancing NK cell cytotoxicity by miR-182 in hepatocellular carcinoma. Hum. Immunol. 77: 667-673. 
  27. Grudzien M, Rapak,A. 2018. Effect of natural compounds on NK cell activation. J. Immunol. Res. 2018: 4868417. 
  28. Wang F, Zhang X, Liu W, Zhou Y, Wei W, Liu D, et al. 2022. Activated natural killer cell promotes nonalcoholic steatohepatitis through mediating JAK/STAT pathway. Cell. Mol. Gastroenterol. Hepatol. 13: 257-274. 
  29. Stangl S, Tontcheva N, Sievert W, Shevtsov M, Niu M, Schmid TE, et al. 2018. Heat shock protein 70 and tumor-infiltrating NK cells as prognostic indicators for patients with squamous cell carcinoma of the head and neck after radiochemotherapy: a multicentre retrospective study of the German Cancer Consortium Radiation Oncology Group (DKTK-ROG). Int. J. Cancer 142: 1911-1925. 
  30. Schantz SP, Campbell BH, Guillamondegui OM. 1986. Pharyngeal carcinoma and natural killer cell activity. Am. J. Surg. 152: 467-474. 
  31. Delahaye NF, Rusakiewicz S, Martins I, Menard C, Roux S, Lyonnet L, et al. 2011. Alternatively spliced NKp30 isoforms affect the prognosis of gastrointestinal stromal tumors. Nat. Med. 17: 700-707. 
  32. DeGregori J, Johnson DG. 2006. Distinct and overlapping roles for E2F family embers in transcription, proliferation and apoptosis. Curr. Mol. Med. 6: 739-748. 
  33. Li Y, Huang J, Yang D, Xiang S, Sun J, Li H, et al. 2018. Expression patterns of E2F transcription factors and their potential prognostic roles in breast cancer. Oncol. Lett. 15: 9216-9230. 
  34. Yang R, Wang M, Zhang G, Bao Y, Wu Y, Li X, et al. 2020. E2F7-EZH2 axis regulates PTEN/AKT/mTOR signalling and glioblastoma progression. Br. J. Cancer 123: 1445-1455. 
  35. Guo H, Zhang L. 2019. MicroRNA-30a suppresses papillary thyroid cancer cell proliferation, migration and invasion by directly targeting E2F7. Exp. Ther. Med. 18: 209-215.