Effect of high glucose on immune escape of pancreatic cancer cells by regulating miR-429 / ZEB1 axis
10.19405/j.cnki.issn1000-1492.2024.07.009
- Author:
Zhichao Zhang
1
;
Guanghui Li
1
;
Xuehe Zhu
1
;
Qiang Wei
1
;
Fei Wang
2
;
Haiping Zhao
2
Author Information
1. Dept of Emergency，Affiliated Hospital of Inner Mongolia Medical University，Hohhot 010010
2. Dept of Hepatobiliary Surgery， Affiliated Hospital of Inner Mongolia Medical University，Hohhot 010010
- Publication Type:Journal Article
- Keywords:
miR-429;
pancreatic cancer;
zinc finger E-box binding homeobox protein 1;
high glucose;
immune es- cape
- From:
Acta Universitatis Medicinalis Anhui
2024;59(7):1166-1174
- CountryChina
- Language:Chinese
-
Abstract:
Objective :To explore the impact of high glucose ( HG) intervention on immune escape of pancreatic cancer cells and its molecular mechanisms．
Methods :PANC-1 cells were treated with different concentrations of glucose (0，7. 5，15，30 mmol / L) for 24 h to establish high glucose intervention PANC-1 cells．miR-429 mimics and its negative control ( mimics NC) were transfected into PANC-1 cells，which were divided into control group，HG group，HG + mimics NC group，HG + mimics group，HG + mimics + oe-NC group，and HG + mimics + oe-ZEB1 group．Flow cytometry was utilized to measure the expression level of cell surface molecule PD-L1 ; qRT-PCR was used to detect the expression levels of miR-429 and ZEB1 mRNA in cells ; Western blot was used to detect the ex- pression level of ZEB1 protein in cells．The above-mentioned PANC-1 cells from each group were co-cultured with CD8 + T cells to establish a co-culture system，and CCK-8 was used to assess cell proliferation activity ; apoptosis levels of cells were measured using flow cytometry ; lactate dehydrogenase ( LDH) release assay was used to detect the killing effect of CD8 + T cells on PANC-1 cells ; dual-luciferase reporter system was used to validate the target- regulatory relationship between miR-429 and ZEB1 .
Results :HG could promote the expression of cell surface mole- cules PD-L1 and ZEB1 in PANC-1 cells (P＜0. 05) ，inhibit the expression of miR-429，and exhibit concentration dependence．Overexpression of miR-429 could significantly suppress the expression of cell surface molecule PD-L1 induced by HG in PANC-1 cells，while overexpression of ZEB1 could reverse the inhibitory effect of miR-429 over- expression on the expression of cell surface molecule PD-L1 induced by HG．After establishing a co-culture system with CD8 + T cells，compared with the control group，the proliferation activity of PANC-1 cells in the HG group sig- nificantly increased，and the apoptosis rate and cytotoxicity significantly decreased (P＜0. 05) ．Compared with the HG + mimics NC group，the proliferation activity of PANC-1 cells in the HG + mimics group significantly decreased， and the apoptosis level and cytotoxicity significantly increased (P＜0. 05) ．Compared with the HG + mimics group， the proliferation activity of PANC-1 cells in the HG + mimics + oe-ZEB1 group significantly increased，and the apop- tosis rate and cytotoxicity significantly decreased (P ＜0. 05 ) ．Dual luciferase reporter gene assay confirmed that miR-429 negatively regulated ZEB1．
Conclusion :High glucose promotes immune escape of PANC-1 cells by down- regulating the expression level of miR-429，negatively regulating the expression of ZEB1 mRNA，and increasing the expression level of cell surface molecule PD-L1 in PANC-1 cells.
- Full text:2024091016475670366高糖通过调控miR-429...对胰腺癌细胞免疫逃逸的影响_张志超.pdf
