[Effect of Silencing FMOD on Proliferation and Migration of H322 Cells and Its Mechanism]
- Sichuan Da Xue Xue Bao Yi Xue Ban. 2020 Mar;51(2):165-170. doi: 10.12182/20200360603.
- 1. Department of Respiratory Medicine, the Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu 322000, China.
Objective: To investigate the regulation of fibromodulin (FMOD) on proliferation, adhesion and migration of Non-Small Cell Lung Cancer cell line H322, and discuss its action mechanism.
Methods: H322 cells were randomly divided into control group, small interfering RNA (siRNA) silencing FMOD ( FMOD siRNA) group and control siRNA (Con siRNA) group. FMOD siRNA and Con siRNA were transfected into H322 cells. The cell viability of each group was detected by CCK-8 method. The adhesion ability of cells was detected by fluorescein diacetate (FDA) fluorescent staining. The cell migration ability was detected by Transwell method. Real time-PCR was used to detect the mRNA expressions of Cyclin D1, intercellular adhesion molecule -1 （ICAM-1）, E-cadherin, FMOD, transforming growth factor-β (TGF-β), SMAD2, SMAD3, SMAD4 and Smad7 in cells. The protein expressions of Cyclin D1, ICAM-1, E-cadherin, FMOD, TGF-β1, SMAD2, SMAD3, SMAD4 and Smad7 were detected by Western blot.
Results: Compared with the Con siRNA group, the cell viability, cell adhesion and migration ability of the FMOD siRNA group were decreased, and the difference was statistically significant ( P<0.01). There was no significant difference between the control group and the Con siRNA group. Real time-PCR and Western blot results showed that the mRNA and protein expression levels of Cyclin D1, ICAM-1, TGF-β1, SMAD2, SMAD3 and SMAD4 were decreased in FMOD siRNA group, compared with Con siRNA group, while the mRNA and protein expression levels of E-cadherin and Smad7 are elevated.
Conclusion: Silencing of the FMOD gene significantly reduces the proliferation, adhesion and migration of H322 cells, which may be conducted by inhibiting the TGF-β/Smad signaling pathway.