Hepatocyte proliferation and apoptosis under regulation of human telomerase reverse transcriptase gene-modified bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2015.32.007

Abstract

Abstract:

BACKGROUND: Studies have shown that human telomerase reverse transcriptase gene (hTERT) transfection can significantly extend the life cycle of bone marrow mesenchymal stem cells so that the cells can continue to maintain pluripotency.
OBJECTIVE: To investigate the effects of hTERT gene-modified bone marrow mesenchymal stem cells on hepatocyte proliferation and apoptosis.
METHODS: Bone marrow mesenchymal stem cells from rats were isolated and cultured using direct adherent method. Then, hTERT eukaryotic expression plasmid, pCIneo-hTERT, was transferred into the cells using liposome transfection method. The hTERT-modified bone marrow mesenchymal stem cells were co-cultured with hepatocytes at 1:1 (observation group), and meanwhile, non-transfected bone marrow mesenchyam stem cells were co-cultured with hepatocytes at 1:1 (control group), and hepatocytes cultured alone served as single culture group. Effects of bone marrow mesenchymal stem cells on hepatocyte proliferation and apoptosis were observed by MTT assay and immunofluorescence staining.
RESULTS AND CONCLUSION: The proliferative rate of hepatocytes was significantly higher in the observation group than the control and single culture groups (P < 0.05), and the survival rate of hepatocytes was significantly higher in the observation group than the single culture group (P < 0.05). Experimental findings suggest hTERT-modified bone marrow mesenchymal stem cells can inhibit hepatocyte apoptosis but promote hepatocyte proliferation, so as to improve hepatocyte function.

Key words: 干细胞, 骨髓干细胞, 人端粒酶反转录酶基因, 骨髓间充质干细胞, 肝细胞, 细胞增殖, 细胞凋亡

CLC Number:

R394.2

Bai Dong, Zhou Zhong-xiao, Zhang Jian. Hepatocyte proliferation and apoptosis under regulation of human telomerase reverse transcriptase gene-modified bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(32): 5118-5122.

Figures/Tables 3

References

[1] 张业伟,牛坚,赵何伟,等.干细胞表面标志物CD90及端粒酶逆转录酶在原发性肝癌中的表达及临床意义[J].中华肝胆外科杂志, 2012,18(1):37-39.

[2] Thenappan A, Li Y, Kitisin K, et al. Role of transforming growth factor beta signaling and expansion of progenitor cells in regenerating liver. Hepatology. 2010;51(4):1373-1382.

[3] 张莉萍,康格非.原代肝细胞培养的研究现状[J].国外医学:临床生物化学与检验学分册,2004,25(3):193-196.

[4] Liang X, So YH, Cui J, et al. The low-dose ionizing radiation stimulates cell proliferation via activation of the MAPK/ERK pathway in rat cultured mesenchymal stem cells. J Radiat Res. 2011;52(3):380-386.

[5] Kisseleva T, Gigante E, Brenner DA. Recent advances in liver stem cell therapy. Curr Opin Gastroenterol. 2010;26(4):395- 402.

[6] 刘京龙,邓志华,郭素雅,等. Ad-hTERTp-单纯疱疹病毒胸苷激酶/丙氧鸟苷系统联合奥沙利铂对人类肝癌细胞的影响[J].肿瘤研究与临床,2011,23(8):526-531.

[7] Crop MJ, Baan CC, Korevaar SS, et al. Human adipose tissue-derived mesenchymal stem cells induce explosive T-cell proliferation. Stem Cells Dev. 2010;19(12):1843-1853.

[8] 何吉文,李华,向国安,等.肿瘤靶向性细胞穿膜肽介导小分子干扰RNA对肝癌细胞的抑制作用[J].中华实验外科杂志,2013,30(10): 2040-2043.

[9] Shi Y, Xia YY, Wang L, et al. Neural cell adhesion molecule modulates mesenchymal stromal cell migration via activation of MAPK/ERK signaling. Exp Cell Res. 2012;318(17): 2257-2267.

[10] Matuskova M, Hlubinova K, Pastorakova A, et al. HSV-tk expressing mesenchymal stem cells exert bystander effect on human glioblastoma cells. ancer Lett. 2010;290(1):58-67.

[11] 周佳美,向慧玲,朱争艳,等.大鼠骨髓间充质干细胞条件培养基对大鼠肝癌细胞系CBRH,7919增殖能力的影响[J].天津医科大学学报,2011,17(4):455-458．

[12] Lim YS. Acute liver failure in Korea: etiology, prognosis and treatment. Korean J Hepatol. 2010;16(1):5-18.

[13] 邵志红,王培军,李铭华,等.大鼠骨髓间充质干细胞移植对Walker-256肝癌生长影响的实验研究[J].中华医学杂志,2009, 89(7):491-496.

[14] Gilchrist ES, Plevris JN. Bone marrow-derived stem cells in liver repair: 10 years down the line. Liver Transpl. 2010;16(2): 118-129.

[15] Aquino JB, Bolontrade MF, García MG, et al. Mesenchymal stem cells as therapeutic tools and gene carriers in liver fibrosis and hepatocellular carcinoma. Gene Ther. 2010; 17(6):692-708.

[16] Bergfeld SA, DeClerck YA. Bone marrow-derived mesenchymal stem cells and the tumor microenvironment. Cancer Metastasis Rev. 2010;29(2):249-261.

[17] Park SA, Ryu CH, Kim SM, et al. CXCR4-transfected human umbilical cord blood-derived mesenchymal stem cells exhibit enhanced migratory capacity toward gliomas. Int J Oncol. 2011; 38(1):97-103..

[18] Li GC, Ye QH, Xue YH, et al. Human mesenchymal stem cells inhibit metastasis of a hepatocellular carcinoma model using the MHCC97-H cell line. Cancer Sci. 2010;101(12):2546- 2553.

[19] 林沪,陈黎明,王福生,等.间充质干细胞在肝脏的分化机制研究进展[J].实用肝脏病杂志,2010,13(3):238-240.

[20] Røsland GV, Svendsen A, Torsvik A, et al. Long-term cultures of bone marrow-derived human mesenchymal stem cells frequently undergo spontaneous malignant transformation. Cancer Res. 2009;69(13):5331-5339.

[21] Hasebe Y, Hasegawa S, Hashimoto N, et al. Analysis of cell characterization using cell surface markers in the dermis. J Dermatol Sci. 2011;62(2):98-106.

[22] 顾劲扬,施晓雷,张悦,等. 骨髓间充质干细胞维持猪肝细胞形态与功能的实验研究[J].中华肝胆外科杂志,2010,16(2):130-133．

[23] Klopp AH, Gupta A, Spaeth E, et al. Concise review: Dissecting a discrepancy in the literature: do mesenchymal stem cells support or suppress tumor growth. Stem Cells. 2011;29(1):11-19.

[24] 沈中,徐小梅,孟琴,等.原代肝细胞体外培养用于利福平和异烟肼肝毒性的研究[J].中国药科大学学报,2005,36(3):250-253.

[25] Zou C, Luo Q, Qin J, et al. Osteopontin promotes mesenchymal stem cell migration and lessens cell stiffness via integrin β1, FAK, and ERK pathways. Cell Biochem Biophys. 2013;65(3):455-462.

[26] Abdel aziz MT, El Asmar MF, Atta HM, et al. Efficacy of mesenchymal stem cells in suppression of hepatocarcinorigenesis in rats: possible role of Wnt signaling. J Exp Clin Cancer Res. 2011;30:49.

[27] Ho IA, Chan KY, Ng WH, et al. Matrix metalloproteinase 1 is necessary for the migration of human bone marrow-derived mesenchymal stem cells toward human glioma. Stem Cells. 2009;27(6):1366-1375.

[28] Han Z, Jing Y, Zhang S, et al. The role of immunosuppression of mesenchymal stem cells in tissue repair and tumor growth. Cell Biosci. 2012;2(1):8.

[29] 王雪峰,梅佳玮,廖传文,等.骨髓间充质干细胞与小鼠肝癌肝移植术后复发关系的实验观察[J].中华医学杂志,2014,94(6): 455-458.

[30] Gao P, Ding Q, Wu Z, et al. Therapeutic potential of human mesenchymal stem cells producing IL-12 in a mouse xenograft model of renal cell carcinoma. Cancer Lett. 2010; 290(2):157-166.