Effects of FoxM1 expression regulated by lentiviral transfection on the proliferation and invasion of human intrahepatic cholangiocarcinoma and the expression of MMPs

doi:10.11958/20220114

Abstract

Abstract:

Objective To investigate the effects of FoxM1 regulated by lentiviral transfection on the proliferation and invasion of human intrahepatic cholangiocarcinoma (ICC) and the expression levels of MMP-9 and MMP-2 in vitro. Methods Western blot assay was used to detect the FoxM1 protein expression levels of ICC cell lines HCCC-9810, RBE and SSP-25. The higher expression level of cell line was selected as the down-regulated FoxM1 cell line, and the lower one was selected as the up-regulated FoxM1 cell line. FoxM1 plasmid and shRNA lentiviral vector transfection targeting up-regulating and down-regulating ICC cell lines were established a cell line that stably up-regulating or down-regulating FoxM1 cells (verified by Western blot assay). MTT method was used to detect cell proliferation after transfection, and Transwell invasion test was used to detect cell invasion. qPCR was used to detect the expression levels of MMP-9 and MMP-2 mRNA in stably transfected cell lines of each group. Results The protein of FoxM1 expression of SSP-25 was the highest, and HCCC-9810 was the lowest. Therefore, SSP-25 was selected as the target cell line for down-regulating FoxM1 expression, and HCCC-9810 was selected as the target cell line for up-regulating FoxM1 expression. The lentiviral transfection successfully constructed stable overexpression (the HCCC-9810-FoxM1 group) and down-regulated FoxM1 cell line (the SSP-25-shFoxM1 group). The proliferation and invasion ability were significantly higher in the HCCC-9810-FoxM1 group than those of the HCCC-9810-Control group (P<0.05). The proliferation and invasion ability were significantly lower in the SSP-25-shFoxM1 group than those of the SSP-25-Control group (P<0.05). In the HCCC-9810-FoxM1 group, the mRNA expression levels of MMP-9 and MMP-2 were significantly higher than those of the HCCC-9810-Control group, while the mRNA expression levels of MMP-9 and MMP-2 were significantly lower in the SSP-25-shFoxM1 group than those of the SSP-25-Control group (P<0.05). Conclusion FoxM1 promotes the proliferation and invasion of ICC cells. FoxM1 may affect the expression levels of MMP-9 and MMP-2. FoxM1 may serve as a potential biomarker of ICC.

Key words: forkhead box protein M1, cholangiocarcinoma, cell proliferation, neoplasm invasiveness, matrix metalloproteinase 9, matrix metalloproteinase 2

CLC Number:

R735.7

Figures/Tables 7

References 16

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基因名称	引物序列（5′→3′）	产物大小（bp）
FoxM1	上游：AAGCCAGGCTGGAAGAACTC	150
FoxM1	下游：CTGCTGCTTAAACACCTGGTC	150
MMP-9	上游：TCGTGGTTCCAACTCGGTTT	189
MMP-9	下游：GGGTGTAGAGTCTCTCGCTG	189
MMP-2	上游：GGACTTAGACCGCTTGGCTT	177
MMP-2	下游：GTGTTCAGGTATTGCATGTGCT	177
GAPDH	上游：CGGGAAGGAAATGAATGGGC	179
GAPDH	下游：GCCCAATACGACCAAATCAGAGAA	179

基因名称	引物序列（5′→3′）	产物大小（bp）
FoxM1	上游：AAGCCAGGCTGGAAGAACTC	150
FoxM1	下游：CTGCTGCTTAAACACCTGGTC	150
MMP-9	上游：TCGTGGTTCCAACTCGGTTT	189
MMP-9	下游：GGGTGTAGAGTCTCTCGCTG	189
MMP-2	上游：GGACTTAGACCGCTTGGCTT	177
MMP-2	下游：GTGTTCAGGTATTGCATGTGCT	177
GAPDH	上游：CGGGAAGGAAATGAATGGGC	179
GAPDH	下游：GCCCAATACGACCAAATCAGAGAA	179

组别	24 h	48 h	72 h
HCCC-9810-Control组	0.36±0.01	0.76±0.07	1.21±0.08
HCCC-9810-FoxM1组	0.34±0.02	1.07±0.10	2.08±0.08
t	1.107	4.544^＊	11.322^＊＊
SSP-25-Control组	0.35±0.01	0.81±0.06	1.65±0.09
SSP-25-shFoxM1组	0.36±0.01	0.59±0.06	1.02±0.09
t	0.725	3.908^＊	7.288^＊＊

组别	24 h	48 h	72 h
HCCC-9810-Control组	0.36±0.01	0.76±0.07	1.21±0.08
HCCC-9810-FoxM1组	0.34±0.02	1.07±0.10	2.08±0.08
t	1.107	4.544^＊	11.322^＊＊
SSP-25-Control组	0.35±0.01	0.81±0.06	1.65±0.09
SSP-25-shFoxM1组	0.36±0.01	0.59±0.06	1.02±0.09
t	0.725	3.908^＊	7.288^＊＊

组别	MMP-9	MMP-2	FoxM1
HCCC-9810-Control组	1.08±0.12	1.00±0.09	1.05±0.11
HCCC-9810-FoxM1组	2.29±0.25	2.40±0.31	2.71±0.28
t	10.464^＊＊	15.640^＊＊	15.044^＊＊
SSP-25-Control组	1.05±0.05	1.04±0.03	1.03±0.04
SSP-25-shFoxM1组	0.47±0.01	0.53±0.08	0.27±0.11
t	10.521^＊＊	15.992^＊＊	18.603^＊＊