Experimental research on the biological behavior of human liver cancer stem cells

doi:10.3877/cma.j.issn.2095-3232.2014.01.011

Abstract

Abstract:

Objective

To study the biological behavior of human liver cancer stem cells.

Methods

Human liver cancer MHCC97H cells in logarithmic growth phase were inoculated in the serum free medium for suspension cultivation and human liver cancer MHCC97H stem cells with stable propagation were acquired. Human liver cancer MHCC97H stem cell microspheres and human liver cancer MHCC97H cells were collected. Messenger ribonucleic acid (mRNA) of cell markers including cluster of differentiation (CD) 133, CD90, epithelial cell adhesion molecule (EpCAM) was detected by reverse transcription polymerase chain reaction (RT-PCR). Colony formation assay, tumor cell invasion assay (Transwell assay) and nude mouse tumorigenicity assay were performed respectively to observe the colony formation, cell invasion and tumorigenicity of two kinds of cells. The experimental data of two kinds of cells were compared by t test or t′ test.

Results

The mean contents of CD133, CD90, EpCAM mRNA in human liver cancer MHCC97H stem cells (32.70±0.20, 66.30±0.32, 115.40±4.00) were significantly higher than those in human liver cancer MHCC97H cells (1.27±0.29, 13.60±1.36, 1.34±0.40) (t′=117.8, 53.97, 83.37; P<0.05). The colony forming efficiency was (213±10)% in human liver cancer MHCC97H stem cells and was (54±11)% in human liver cancer MHCC97H cells, where significant difference was observed (t=13.11, P<0.05). Through the Transwell assay, the membrane permeating cell count was (587±120) /visual fields in human liver cancer MHCC97H stem cells, and was (97±13) /visual fields in human liver cancer MHCC97H cells, where significant difference was observed (t=6.38, P<0.05). In the nude mouse tumorigenicity assay, subcutaneous transplanted tumors were observed in nude mouse 4 weeks after inoculating 2×10³ human liver cancer MHCC97H stem cells and the tumor formation rate was 1/6. Subcutaneous transplanted tumors were observed in all nude mice by inoculating 2×10⁵ human liver cancer MHCC97H stem cells, and the tumor formation rate was 6/6. Subcutaneous transplanted tumors were observed in nude mice by inoculating at least 2×10⁵ human liver cancer MHCC97H cells, and the tumor formation rate was 2/6.

Conclusion

Compared with the human liver cancer MHCC97H cells, human liver cancer MHCC97H stem cells possess stronger invasion and higher tumor formation rate.

Key words: Liver neoplasms, experimental, Stem cells, Culture media, serum-free, Microspheres

Nan Jiang, Danyun Ruan, Yang Li, Guoying Wang, Binsheng Fu, Genshu Wang, Hua Li, Jian Zhang, Yang Yang, Guihua Chen. Experimental research on the biological behavior of human liver cancer stem cells[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2014, 03(01): 41-45.

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References 14

[1]	Clarke MF,Dick JE,Dirks PB, et al. Cancer stem cells: perspectives on current status and future directionsAACR Workshop on cancer stem cells[J].. Cancer Res, 2006, 66(19): 9339-9344.
[2]	Reya T,Morrison SJ,Clarke MF, et al. Stem cells, cancer, and cancer stem cells[J]. Nature, 2001, 414(6859): 105-111.
[3]	Al-Hajj M,Wicha MS,Benito-Hernandez A, et al. Prospective identification of tumorigenic breast cancer cells[J]. Proc Natl Acad Sci U S A, 2003, 100(7): 3983-3988.
[4]	Singh SK,Hawkins C,Clarke ID, et al. Identification of human brain tumour initiating cells[J]. Nature, 2004, 432(7015): 396-401.
[5]	Martens DJ,Tropepe V,van der Kooy D. Separate proliferation kinetics of fibroblast growth factor-responsive and epidermal growth factor-responsive neural stem cells within the embryonic forebrain germinal zone[J]. J Neurosci, 2000, 20(3): 1085-1095.
[6]	Singh SK,Clarke ID,Terasaki M, et al. Identification of a cancer stem cell in human brain tumors[J]. Cancer Res, 2003, 63(18): 5821-5828.
[7]	Ponti D,Costa A,Zaffaroni N, et al. Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties[J]. Cancer Res, 2005, 65(13): 5506-5511.
[8]	Hong-mei P,Gui-an C. Serum-free medium cultivation to improve efficacy in establishment of human embryonic stem cell lines[J]. Hum Reprod, 2006, 21(1): 217-222.
[9]	Hemmati HD,Nakano I,Lazareff JA, et al. Cancerous stem cell can arise from pediatric brain tumors[J]. Proc Natl Acad Sci U S A, 2003, 100(25): 15178-15183.
[10]	Reynolds BA,Weiss S. Clonal and population analyses demonstrate that an EGF-responsive mammalian embryonic CNS precursor is a stem cell[J]. Dev Biol, 1996, 175(1): 1-13.
[11]	Haraguchi N,Utsunomiya T,Inoue H, et al. Characterization of a side population of cancer cells from human gastrointestinal system[J]. Stem Cells, 2006, 24(3): 506-513.
[12]	Yin S,Li J,Hu C, et al. CD133 positive hepatocellular carcinoma cells possess high capacity for tumorigenicity[J]. Int J Cancer, 2007, 120(7): 1444-1450.
[13]	Yang ZF,Ho DW,Ng MN, et al. Significance of CD90⁺ cancer stem cells in human liver cancer[J]. Cancer Cell, 2008, 13(2): 153-166.
[14]	Yamashita T,Ji J,Budhu A, et al. EpCAM-positive hepatocellular carcinoma cells are tumor-initiating cells with stem/progenitor cell features[J]. Gastroenterology, 2009, 136(3): 1012-1024.

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