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CLC number: R762

On-line Access: 2012-12-07

Received: 2012-02-23

Revision Accepted: 2012-07-02

Crosschecked: 2012-10-31

Cited: 8

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Journal of Zhejiang University SCIENCE B 2012 Vol.13 No.12 P.997-1005


Effect of mitomycin on normal dermal fibroblast and HaCat cell: an in vitro study

Author(s):  Yao-wen Wang, Ji-hao Ren, Kun Xia, Shu-hui Wang, Tuan-fang Yin, Ding-hua Xie, Li-hua Li

Affiliation(s):  Otolaryngology-Head and Neck Surgery Department, Ningbo First Hospital, Ningbo 315000, China; more

Corresponding email(s):   jihao5114@sina.com

Key Words:  Mitomycin, Fibroblast, HaCat cell, Apoptosis, TGF-β, 1, bFGF, Procollagen

Yao-wen Wang, Ji-hao Ren, Kun Xia, Shu-hui Wang, Tuan-fang Yin, Ding-hua Xie, Li-hua Li. Effect of mitomycin on normal dermal fibroblast and HaCat cell: an in vitro study[J]. Journal of Zhejiang University Science B, 2012, 13(12): 997-1005.

@article{title="Effect of mitomycin on normal dermal fibroblast and HaCat cell: an in vitro study",
author="Yao-wen Wang, Ji-hao Ren, Kun Xia, Shu-hui Wang, Tuan-fang Yin, Ding-hua Xie, Li-hua Li",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Effect of mitomycin on normal dermal fibroblast and HaCat cell: an in vitro study
%A Yao-wen Wang
%A Ji-hao Ren
%A Kun Xia
%A Shu-hui Wang
%A Tuan-fang Yin
%A Ding-hua Xie
%A Li-hua Li
%J Journal of Zhejiang University SCIENCE B
%V 13
%N 12
%P 997-1005
%@ 1673-1581
%D 2012
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1200055

T1 - Effect of mitomycin on normal dermal fibroblast and HaCat cell: an in vitro study
A1 - Yao-wen Wang
A1 - Ji-hao Ren
A1 - Kun Xia
A1 - Shu-hui Wang
A1 - Tuan-fang Yin
A1 - Ding-hua Xie
A1 - Li-hua Li
J0 - Journal of Zhejiang University Science B
VL - 13
IS - 12
SP - 997
EP - 1005
%@ 1673-1581
Y1 - 2012
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1200055

Objective: To evaluate the effects of mitomycin on the growth of human dermal fibroblast and immortalized human keratinocyte line (haCat cell), particularly the effect of mitomycin on intracellular messenger RNA (mRNA) synthesis of collagen and growth factors of fibroblast. Methods: The normal dermal fibroblast and haCat cell were cultured in vitro. Cell cultures were exposed to 0.4 and 0.04 mg/ml of mitomycin solution, and serum-free culture medium was used as control. The cellular morphology change, growth characteristics, cell proliferation, and apoptosis were observed at different intervals. For the fibroblasts, the mRNA expression changes of transforming growth factor (TGF)-β1, basic fibroblast growth factor (bFGF), procollagen I, and III were detected by reverse transcription polymerase chain reaction (RT-PCR). Results: The cultured normal human skin fibroblast and haCat cell grew exponentially. A 5-min exposure to mitomycin at either 0.4 or 0.04 mg/ml caused marked dose-dependent cell proliferation inhibition on both fibroblasts and haCat cells. Cell morphology changed, cell density decreased, and the growth curves were without an exponential phase. The fibroblast proliferated on the 5th day after the 5-min exposure of mitomycin at 0.04 mg/ml. Meanwhile, 5-min application of mitomycin at either 0.04 or 0.4 mg/ml induced fibroblast apoptosis but not necrosis. The apoptosis rate of the fibroblast increased with a higher concentration of mytomycin (p<0.05). A 5-min exposure to mitomycin at 0.4 mg/ml resulted in a marked decrease in the mRNA production of TGF-β;1, procollagen I and III, and a marked increase in the mRNA production of bFGF. Conclusions: mitomycin can inhibit fibroblast proliferation, induce fibroblast apoptosis, and regulate intracellular protein expression on mRNA levels. In addition, mitomycin can inhibit haCat cell proliferation, so epithelial cell needs more protecting to avoid mitomycin 19;s side effect when it is applied clinically.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Bank, H.L., 1988. Rapid assessment of islet viability with acridine orange and propidium iodide. In Vitro Cel. Dev. Biol., 24(4):266-269.

[2]Banthia, V., Selesnick, S.H., 2003. Mitomycin-C in the Post Surgical Ear Canal. American Academy of Otolaryngology/Head and Neck Surgery. Annual Meeting, Orlando, Florida, USA, p.882-886.

[3]Battelino, S., Hocevar-Boltezar, I., Zargi, M., 2005. Intraoperative use of mitomycin C in fibrous atresia of the external auditory canal. Ear Nose Throat J., 84(12):776-779.

[4]Chen, T., Kunnavatana, S.S., Koch, R.J., 2006. Effects of mitomycin-C on normal dermal fibroblasts. Laryngoscope, 116(4):514-517.

[5]Crowston, J.G., Chang, L.H., Constable, P.H., Daniels, J.T., Akhar, A.N., Khaw, P.T., 2002. Apoptosis gene expression and death receptor signaling in mitomycin C-treated human Tenon capsule fibroblasts. Invest. Ophthalmol. Vis. Sci., 43(3):692-699.

[6]Ferguson, B., Gray, S.D., Thibeault, S., 2005. Time and dose effects of mitomycin C on extracellular matrix fibroblasts and proteins. Laryngoscope, 115(1):110-115.

[7]Gospodarowicz, D., 1991. Biological activities of fibroblast growth factors. Ann. N. Y. Acad. Sci., 638:1-8.

[8]Gray, S.D., Tritle, N., Li, W., 2003. The effect of mitomycin on extracellular matrix proteins in a rat wound model. Laryngoscope, 113(2):237-242.

[9]Hong, R., Lum, J., Koch, R.J., 1999. Growth of keloid-producing fibroblasts in commercially available serum-free media: a comparative study. Otolaryngol. Head Neck Surg., 121(4):469-473.

[10]Kikuchi, K., Kakano, T., Takehara, K., 1995. Effects of various growth factors and histamine on cultured keloid fibroblasts. Dermatology, 190(1):4-8.

[11]Kim, T.I., Tchah, H., Lee, S., Cho, B.J., Kook, M.S., 2003. Apoptosis in keratocytes caused by mitomycin C. Invest. Ophthalmol. Vis. Sci., 44(5):1912-1917.

[12]Kim, T.I., Tchah, H., Cho, E.H., Kook, M.S., 2004. Evaluation of safety of cultured corneal fibroblasts with cotreatment of alcohol and mitomycin C. Invest. Ophthalmol. Vis. Sci., 45(1):86-92.

[13]Moses, H.L., Yang, E.Y., Peitenpol, J.A., 1990. TGF-β stimulation and inhibition of cell proliferation: new mechanistic insights. Cell, 63(2):245-257.

[14]Occleston, N.L., Daniels, J.T., Tarnuzzer, R.W., Sethi, K.K., Alexander, R.A., Bhattacharya, S.S., Schultz, G.S., Khaw, P.T., 1997. Single exposures to antiproliferatives: long-term effects on ocular fibroblasts wound-healing behavior. Invest. Ophthalmol. Vis. Sci., 38(10):1998-2007.

[15]Peltonen, J., Hsiao, L.L., Jaakkola, S., Sollberg, S., Aumailley, M., Timpl, R., Chu, M.L., Uitto, J., 1991. Activation of collagen gene expression in keloids: co-localization of type I and VI collagen and transforming growth factor-β1 mRNA. J. Invest. Dermatol., 97(2):240-248.

[16]Prasad, M., Ward, R.F., April, M.M., Bent, J.P., Froehlich, P., 2002. Topical mitomycin as an adjunct to choanal atresia repair. Arch. Otolaryngol. Head Neck Surg., 128(4):398-400.

[17]Ragab, S., 2005. The effect of radiofrequency and mitomycin C on the closure rate of human. tympanostomy. Otol. Neurotol., 26(3):355-360.

[18]Rahbar, R., Jones, D.T., Nuss, R.C., Roberson, D.W., Kenna, M.A., McGill, T.J., Healy, G.B., 2002. The role of mitomycin in the prevention and treatment of scar formation in the pediatric aerodigestive tract. Arch. Otolaryngol. Head Neck Surg., 128(4):401-406.

[19]Schwartzman, R.A., Cidlowski, J.A., 1993. Apoptosis: the biochemistry and molecular biology of programmed cell death. Endocr. Rev., 14(2):133-151.

[20]Seong, G.J., Park, C., Kim, C.Y., Hong, Y.J., So, H.S., Kim, S.D., Park, R., 2005. Mitomycin-C induced the apoptosis of human Tenon’s capsule fibroblast by activation of c-Jun N-terminal kinase 1 and caspase-3 protease. Invest. Ophthalmol. Vis. Sci., 46(10):3545-3552.

[21]Simman, R., Hashim, A., Frances, W., 2003. Effect of mitomycin-C on keloid fibroblasts: an in vitro study. Ann. Plast. Surg., 50(1):71-76.

[22]Simpson, C.B., James, J.C., 2006. The efficacy of mitomycin-C in the treatment of laryngotracheal-stenosis. Laryngoscope, 116(10):1923-1925.

[23]Stewart, C.E.IV, Kim, J.Y., 2006. Application of mitomycin-C for head and neck keloids. Otolaryngol. Head Neck Surg., 135(6):946-950.

[24]Tahery, M.M., Lee, D.A., 1989. Review: pharmacologic control of wound healing in glaucoma filtration surgery. J. Ocul. Pharmacol., 5(2):155-179.

[25]Younai, S., Nichter, L.S., Wellisz, T., Reinisch, J., Nimni, M.E., Tuan, T.L., 1994. Modulation of collagen synthesis by transforming growth factor-beta in keloid and hypertrophic scar fibroblasts. Ann. Plast. Surg., 33(2):148-151.

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