Culture of Malignant Pleural Mesothelioma Cells and the Effects of CDKN2B on Cancer Cell
10.12259/j.issn.2095-610X.S20240105
- VernacularTitle:恶性胸膜间皮瘤细胞培养条件及CDKN2B对癌细胞的作用
- Author:
Xiaochuan YIN
1
;
Ruiyang YIN
;
Ranhua LI
;
Fangqi CAI
;
Yue CUI
;
Tao BI
;
Xinghe TONG
Author Information
1. 昆明医科大学第一附属医院胸外科,云南 昆明 650032
- Keywords:
Malignant pleural mesothelioma;
Culture conditions;
RPMI-1640 culture medium;
CDKN2B;
Proliferation;
Invasion
- From:
Journal of Kunming Medical University
2024;45(1):28-34
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of different culture conditions(RPMI-1640,DMEM and DMEM/F12 medium)on the passage of MPM cells isolated from the tissues of Malignant pleural mesothelioma(MPM),and to study the effects of CDKN2B on the proliferation,invasion and apoptosis of MPM cells.Methods MPM cells were isolated from MPM tissues and cultured in RPMI-1640,DMEM and DMEM/F12 medium,respectively.Cell proliferation was examined by CCK-8,and the nuclei and chromosomes were observed by Wright-Giemsa staining.Fluorescence intensities of Calretinin,CD141,CK5,EMA and WT-1 were conducted by immunofluorescence assay.The mRNA and protein expression of CDKN2B were detected by RT-qPCR and Western blot,respectively.Transwell was used to detect cell invasion and flow cytometry was used to detect cell apoptosis.Results The established MPM cells showed good viability when passaged to the 10th generation in RPMI-1640,DMEM and DMEM/F12 cultures,and the MPM markers Calretinin,CD141,CK5,EMA and WT-1 were all expressed in the cells.The viability of MPM cells in RPMI-1640 culture medium was relatively stable.CDKN2B was downregulated in MPM cells(P<0.05),and overexpression of CDKN2B significantly suppressed the proliferation(P<0.05),invasion(P<0.05)and epithelial interstitial transformation of MPM cells(P<0.01),and promoted the apoptosis(P<0.01).Conclusion The established MPM cells were stably passaged in RPMI-1640 culture medium,and CDKN2B may be a potential target for the diagnosis and treatment of MPM.
