Effect of liver cancer derived mesenchymal stem cell on invasion of liver cancer cells

doi:10.3877/cma.j.issn.2095-3232.2016.06.014

Abstract

Abstract:

Objective

To investigate the effect and mechanism of liver cancer derived mesenchymal stem cell (LCMSC) on the invasion of liver cancer cells.

Methods

The expressions of interleukin (IL)-6, IL-8 and chemotactic factors CXCL1, CXCL5 and CXCL12 mRNA in the bone marrow derived mesenchymal stem cell (BMSC) and LCMSC were detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of protein CXCL12 in the supernate of LCMSC was detected by enzyme-linked immunosorbent assy (ELISA) and Western blot. Different cells were co-cultured and divided into the HepG2+BMSC, HepG2+LCMSC and HepG2+LCMSC+siRNA-CXCL12 groups. The effect of CXCL12 on the invasion of liver cancer HepG2 cells were detected by Transwell migration assay. The experiment data were compared using one way analysis of variance and LSD-t test or t test.

Results

The expression of CXCL12 mRNA in LCMSC was 60.3±2.4, significantly higher than 13.8±1.8 in BMSC (t=15.68, P<0.05). The expression of protein CXCL12 in the supernate of LCMSC was (31.5±1.7) ng/L, significantly higher than (14.3±1.5) ng/L in BMSC (t=7.60, P<0.05). And the expression of protein CXCL12 was up-regulated. Transwell migration assay indicated that the quantity of membrane-invasion cells in the HepG2+LCMSC group was 110±12, significantly higher than 65±9 in the HepG2+BMSC group and 76±7 in the HepG2+LCMSC+siRNA-CXCL12 group (LSD-t=5.25, 4.86; P<0.05).

Conclusion

CXCL12 is highly expressed in LCMSC. LCMSC may enhance the invasion of HepG2 cells through up-regulating the expression of CXCL12. The invasion of liver cancer cells can be effectively weakend by silencing the CXCL12 gene with siRNA.

Key words: Mesenchymal stem cells, Carcinoma, hepatocellular, Neoplasm invasiveness, Chemokine CXCL12

Zhicheng Yao, Jiezhong Wu, Zhiyong Xiong, Jianliang Xu, Meihai Deng, Heping Fang. Effect of liver cancer derived mesenchymal stem cell on invasion of liver cancer cells[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2016, 05(06): 404-408.

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References 21

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基因	引物序列
IL-6	F:5'-CCAGTACCCCCAGGAGAAGAT-3'
IL-6	R:5'-TTGCCTTTTTCTGCAGGAAC-3'
IL-8	F:5'-TTGGCAGCCTTCCTGATTT-3'
IL-8	R:5'-TCAAAAACTTCTCCACAACCC-3'
CXCL1	F:5'-CACCCAAACCGAAGTCATAG-3'
CXCL1	R:5'-AAGCCAGCGTTCACCAGA-3'
CXCL5	F:5'-CGCTGTCATCCTCATTGCTA-3'
CXCL5	R:5'-GAGCACTTGCCACTGGTGTA-3'
CXCL12	F:5'-AGTCAGGTGGTGGCTTAACAG-3'
CXCL12	R:5'-AGAGGAGGTGAAGGCAGTGG-3'
GAPDH	F:5'-ACCACAGTCCATGCCATCAC-3'
GAPDH	R:5'-TCCACCACCCTGTTGCTGTA-3'

基因	引物序列
IL-6	F:5'-CCAGTACCCCCAGGAGAAGAT-3'
IL-6	R:5'-TTGCCTTTTTCTGCAGGAAC-3'
IL-8	F:5'-TTGGCAGCCTTCCTGATTT-3'
IL-8	R:5'-TCAAAAACTTCTCCACAACCC-3'
CXCL1	F:5'-CACCCAAACCGAAGTCATAG-3'
CXCL1	R:5'-AAGCCAGCGTTCACCAGA-3'
CXCL5	F:5'-CGCTGTCATCCTCATTGCTA-3'
CXCL5	R:5'-GAGCACTTGCCACTGGTGTA-3'
CXCL12	F:5'-AGTCAGGTGGTGGCTTAACAG-3'
CXCL12	R:5'-AGAGGAGGTGAAGGCAGTGG-3'
GAPDH	F:5'-ACCACAGTCCATGCCATCAC-3'
GAPDH	R:5'-TCCACCACCCTGTTGCTGTA-3'

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