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小鼠乳腺上皮细胞培养体系的建立及Heregulin-α的调控作用

DOI: 10.1360/052011-26, PP. 296-303

Keywords: 小鼠,乳腺上皮细胞,细胞培养体系,Heregulin-α,生物学特性

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Abstract:

旨在建立完善的小鼠乳腺上皮细胞体外培养体系,探讨Heregulin-α(HRG-α)对该体系乳腺上皮细胞增殖及代谢的影响,为进一步深入研究乳腺上皮细胞的形态、结构和功能,揭示HRG-α在小鼠乳腺发育中的作用及其规律提供理论依据.通过比较不同pH和不同血清浓度配比的生长培养液对小鼠乳腺细胞生长的影响,得出pH7.4,添加10%血清的生长培养液为乳腺上皮细胞生长的最佳条件.通过运用相差显微镜技术、MTT等研究方法进行不同取材时期乳腺上皮细胞的形态、接种存活率、倍增时间、生长曲线等生物学特性比较,获得妊娠15天为较好的乳腺细胞取材时期.采用免疫组化和RT-PCR方法对细胞重要的标志蛋白角蛋白-18进行检测,鉴定所获得的细胞为乳腺上皮细胞,成功建立小鼠乳腺上皮细胞体外培养体系.利用所建立的小鼠乳腺上皮细胞体外培养体系,采用液相色谱技术及MTT方法,初步探讨了HRG-α对小鼠乳腺上皮细胞的影响.结果表明,适宜量HRG-α对小鼠乳腺上皮细胞生长和分泌总蛋白、乳糖均有显著促进作用,0.1~20ng/mL的HRG-α可促进小鼠乳腺上皮细胞的增殖和总蛋白、乳糖含量的增加,20ng/mL时达到最大值(P<0.01),50和100ng/mLHRG-α抑制小鼠乳腺上皮细胞的增殖,降低小鼠乳腺上皮细胞的总蛋白、乳糖含量(P<0.05).

References

[1]  1 李健, 李庆章. 小鼠乳腺发育、泌乳及退化的形态学研究. II. 小鼠乳腺发育的超微形态学研究. 东北农业大学学报, 2007, 38:374-379
[2]  2 耿丽晶, 李庆章. Heregulin-α及其受体在小鼠乳腺中的表达与作用.中国科学: 生命科学, 2009, 39: 362-371
[3]  3 耿丽晶, 李庆章. Heregulin-α促进小鼠妊娠期乳腺上皮细胞的增殖/分化及β酪蛋白的分泌. 中国生物化学与分子生物学报, 2008, 24:50-56
[4]  4 Jones F E, Jerry D J, Guarino B C, et al. Heregulin induces in vivo proliferation and differentiation of mammary epithelium into secretorylobuloalveoli. Cell Growth Differ, 1996, 7: 1031-1038
[5]  5 Hennighausen L, Robinson G W. Think globally act locally: the making of a mouse mammary gland. Genes Dev, 1998, 12: 449-455??
[6]  6 Marte B M, Jeschke M, Graus-Porta D, et al. Neu differentiation factor/heregulin modulates growth and differentiation of HC11 mammaryepithelial cells. Mol Endocrinol, 1995, 9: 14-23??
[7]  7 张以涛, 李庆章. 小白鼠乳腺上皮细胞的分离, 培养及形态学变化. 黑龙江畜牧兽医, 2008, 4: 11-13
[8]  8 Zhu H, Tamot B, Quinton M, et al. Influence of tissue origins and external microenvironment on porcine foetal fibroblast growth,proliferative life span and genome stability. Cell Prolif, 2004, 37: 255-266??
[9]  9 Chen S L, Ren G C, Sha Z X, et al. Establishment of a continuous embryonic cell line from Japanese flounder Paralichthys olivaceus forvirus isolation. Dis Aquat Organ, 2004, 60: 241-246??
[10]  10 Lasfargues E Y, Moore D H, Murray M R. Maintenance of the milk factor in cultures of mouse mammary epithelium. Cancer Res, 1958, 18:1281-1285
[11]  11 Woodward, Jian W X, James L, et al. Proliferation of mouse mammary epithelial cells in vitro: interactions among epidermal growth factor,insulin-like growth factor I, ovarian hormones, and extracellular matrix proteins. The Endocrine Society, 2000, 141: 3578-3586
[12]  12 Pickett P B, Pitelka D R, Hamamoto S T, et al. Occluding junctions and cell behaveior in primary cultures of normal and neoplasticmammary gland cells. Cell Biol, 1975, 66: 316-332??
[13]  13 Medina D, Oborn C J, Kittrell F S, et al. Properties of mouse mammary epithelial cell lines characterized by in vivo transplantation and invitro immunocytochemical methods. JNCI, 1986, 76: 1143-1156
[14]  14 Stingl J, Eaves C J, Zandieh I, et al. Characterization of bipotent mammary epithelial progenitor cells in normal adult human breast tissue.Breast Cancer Res Treat, 2001, 67: 93-109??
[15]  15 Sun Y L, Lin C S, Chou Y C. Establishment and characterization of a spontaneously immortalized porcine mammary epithelial cell line.Cell Biol Int, 2006, 30: 970-976??
[16]  16 Li L, Cleary S, Mandarano M A, et al. The breast proto-oncogene, HRG-alpha regulates epithelial proliferation and lobuloalveolardevelopment in the mouse mammary gland. Oncogene, 2002, 21: 4900-4907??

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