Culture of Malignant Pleural Mesothelioma Cells and the Effects of CDKN2B on Cancer Cell

Xiaochuan YIN; Ruiyang YIN; Ranhua LI; Fangqi CAI; Yue CUI; Tao BI; Xinghe TONG

doi:10.12259/j.issn.2095-610X.S20240105

Volume 45 Issue 1

Jan. 2024

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Article Navigation > Journal of Kunming Medical University > 2024 > 45(1): 28-34

Xiaochuan YIN, Ruiyang YIN, Ranhua LI, Fangqi CAI, Yue CUI, Tao BI, Xinghe TONG. Culture of Malignant Pleural Mesothelioma Cells and the Effects of CDKN2B on Cancer Cell[J]. Journal of Kunming Medical University, 2024, 45(1): 28-34. doi: 10.12259/j.issn.2095-610X.S20240105

Citation:

Xiaochuan YIN, Ruiyang YIN, Ranhua LI, Fangqi CAI, Yue CUI, Tao BI, Xinghe TONG. Culture of Malignant Pleural Mesothelioma Cells and the Effects of CDKN2B on Cancer Cell[J]. Journal of Kunming Medical University, 2024, 45(1): 28-34. doi: 10.12259/j.issn.2095-610X.S20240105

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Culture of Malignant Pleural Mesothelioma Cells and the Effects of CDKN2B on Cancer Cell

doi: 10.12259/j.issn.2095-610X.S20240105

1.
Dept. of Thoracic Surgery，The 1st Affiliated Hospital of Kunming Medical University，Kunming Yunnan 650032
2.
The 2nd Clinical Medicine Department，Kunming Medical University HaiYuan College，Kunming Yunnan 651700，China

Received Date: 2023-10-09
Available Online: 2023-12-19
Publish Date: 2024-01-25

Abstract

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.
- Malignant pleural mesothelioma,
- Culture conditions,
- RPMI-1640 culture medium,
- CDKN2B,
- Proliferation,
- Invasion

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