title: Effect of EBI3 on radiation-induced immunosuppression of cervical cancer HeLa cells by regulating Treg cells through PD-1/PD-L1 pathway.
abstract: This study aimed to investigate the effect of EBI3 on radiation-induced immunosuppression of cervical cancer HeLa cells by regulating Treg cells through PD-1/PD-L1 signaling pathway. A total of 43 adult female Wistar rats were selected and injected with HeLa cells in the caudal vein to construct a rat model of cervical cancer. All model rats were randomly divided into the radiotherapy group ( n = 31) and the control group ( n = 12). The immunophenotype of Treg cells was detected by the flow cytometry. The protein expressions of EBI3, PD-1, and PD-L1 in cervical cancer tissues were tested by the streptavidin-peroxidase method. HeLa cells in the logarithmic growth phase were divided into four groups: the blank, the negative control group, the EBI3 mimics group, and the EBI3 inhibitors group. Western blotting was used to detect PD-1 and PD-L1 protein expressions. MTT assay was performed to measure the proliferation of Treg cells. Flow cytometry was used to detect cell cycle and apoptosis, and CD4+/CD8+T cell ratio in each group. Compared with before and 1 week after radiotherapy, the percentages of CD4+T cells and CD8+T cells were significantly decreased in the radiotherapy group at 1 month after radiotherapy. Furthermore, down-regulation of EBI3 and up-regulation of PD-1 and PD-L1 were observed in cervical cancer tissues at 1 month after radiotherapy. In comparison to the blank and negative control groups, increased expression of EBI3 and decreased expressions of PD-1 and PD-L1 were found in the EBI3 mimics group. However, the EBI3 inhibitors group had a lower expression of EBI3 and higher expressions of PD-1 and PD-L1 than those in the blank and negative control groups. The EBI3 mimics group showed an increase in the optical density value (0.43 ± 0.05), while a decrease in the optical density value (0.31 ± 0.02) was found in the EBI3 inhibitors group. Moreover, compared with the blank and negative control groups, the apoptosis rates of Treg/CD4+T/CD8+T cells were decreased in the EBI3 mimics group, but the EBI3 inhibitors group exhibited an increase in apoptosis rate. In conclusion, over-expression of EBI3 could reduce the apoptosis of Treg/CD4+T/CD8+T cells and prevent radiation-induced immunosuppression of cervical cancer HeLa cells by inhibiting the activation of PD-1/PD-L1 signaling pathway.
authors.affiliation: 1 Cancer Centre, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, P.R. China.
authors.forename: Song-An
authors.initials: SA
authors.lastname: Zhang
authors.affiliation: 1 Cancer Centre, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, P.R. China.
authors.forename: Hu-Er-Xi-Dan
authors.initials: HE
authors.lastname: Niyazi
authors.affiliation: 2 Anus-Intestines Department, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, P.R. China.
authors.forename: Wen
authors.initials: W
authors.lastname: Hong
authors.affiliation: 3 Xinjiang Medical University, Urumqi, P.R. China.
authors.forename: Gu-Li-Xian
authors.initials: GL
authors.lastname: Tuluwengjiang
authors.affiliation: 1 Cancer Centre, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, P.R. China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.affiliation: 1 Cancer Centre, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, P.R. China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: 1 Cancer Centre, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, P.R. China.
authors.forename: Wei-Peng
authors.initials: WP
authors.lastname: Su
authors.affiliation: 1 Cancer Centre, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, P.R. China.
authors.forename: Yong-Xing
authors.initials: YX
authors.lastname: Bao
mesh_terms: D000818:Animals; D060890:B7-H1 Antigen / Q000096:biosynthesis* / Q000235:genetics; D049109:Cell Proliferation / Q000528:radiation effects; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000528:radiation effects; D006367:HeLa Cells; D006801:Humans; D016130:Immunophenotyping; D007165:Immunosuppression Therapy; D007378:Interleukins / Q000096:biosynthesis* / Q000235:genetics; D015778:Minor Histocompatibility Antigens / Q000096:biosynthesis* / Q000235:genetics; D009374:Neoplasms, Experimental / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000532:radiotherapy*; D061026:Programmed Cell Death 1 Receptor / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology / Q000528:radiation effects; D050378:T-Lymphocytes, Regulatory / Q000276:immunology / Q000528:radiation effects; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000532:radiotherapy*
chemical_list: D060890:B7-H1 Antigen; C423236:CD274 protein, human; C097732:EBI3 protein, human; D007378:Interleukins; D015778:Minor Histocompatibility Antigens; C105992:PDCD1 protein, human; D061026:Programmed Cell Death 1 Receptor
keywords: EBI3; PD-1/PD-L1 pathway; Treg cell; cervical cancer; immunosuppression