Spontaneous transformation of human granulosa cell tumours into an aggressive phenotype: a metastasis model cell line
Spontaneous transformation of human granulosa cell tumours into an aggressive phenotype: a metastasis model cell line
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DOI：
10.1186/1471-2407-8-319
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发表时间：
2008-11-04
期刊：
影响因子：
3.8
通讯作者：
Takahashi, Yuji
中科院分区：
文献类型：
--
作者：
Imai, Misa;Muraki, Miho;Takahashi, Yuji
背景：颗粒细胞瘤（GCT）常见于更年期女性，且相对惰性。尽管正常颗粒细胞的生理特性已被广泛研究，但对 GCT 进展的分子机制知之甚少。在这里，我们表征了颗粒肿瘤细胞系 KGN 细胞的独特行为特性，用于 GCT 进展的分子分析。方法：进行群体倍增以检查 KGN 细胞的增殖能力。此外，使用体外侵袭试验测定了这些细胞的侵袭能力。然后通过Western blot分析测定不同代数的KGN细胞中肿瘤标志物的表达水平。最后，在裸鼠皮下注射3个月后观察KGN细胞的生长和转移情况。结果：在体外培养过程中，群体倍增实验显示，晚期传代KGN细胞的生长速度比早期传代细胞快2倍。此外，我们发现高级传代细胞的侵袭性是早期传代细胞的两倍。 p53、骨桥蛋白、BAX 和 BAG-1 等肿瘤标志物的表达模式支持这样的观点：随着传代，KGN 细胞变得更具攻击性。引人注目的是，皮下注射时，早期和晚期传代的 KGN 细胞均转移至肠道。进入裸鼠。此外，当植入高级传代细胞时，会形成更多的肿瘤结节。结论：体外培养的KGN细胞获得了侵袭性表型，细胞活性和生物标志物表达分析证实了这一点。有趣的是，皮下注射 KGN 细胞。是转移性的，结节形成主要发生在肠道中。因此，该细胞系是分析 GCT 进展和体内转移机制的良好模型。
Background: Granulosa cell tumours (GCTs) are frequently seen in menopausal women and are relatively indolent. Although the physiological properties of normal granulosa cells have been studied extensively, little is known about the molecular mechanism of GCT progression. Here, we characterise the unique behavioural properties of a granulosa tumour cell line, KGN cells, for the molecular analysis of GCT progression.Methods: Population doubling was carried out to examine the proliferation capacity of KGN cells. Moreover, the invasive capacity of these cells was determined using the in vitro invasion assay. The expression level of tumour markers in KGN cells at different passages was then determined by Western blot analysis. Finally, the growth and metastasis of KGN cells injected subcutaneously (s.c.) into nude mice was observed 3 months after injection.Results: During in vitro culture, the advanced passage KGN cells grew 2-fold faster than the early passage cells, as determined by the population doubling assay. Moreover, we found that the advanced passage cells were 2-fold more invasive than the early passage cells. The expression pattern of tumour markers, such as p53, osteopontin, BAX and BAG-1, supported the notion that with passage, KGN cells became more aggressive. Strikingly, KGN cells at both early and advanced passages metastasized to the bowel when injected s.c. into nude mice. In addition, more tumour nodules were formed when the advanced passage cells were implanted.Conclusion: KGN cells cultured in vitro acquire an aggressive phenotype, which was confirmed by the analysis of cellular activities and the expression of biomarkers. Interestingly, KGN cells injected s.c. are metastatic with nodule formation occurring mostly in the bowel. Thus, this cell line is a good model for analysing GCT progression and the mechanism of metastasis in vivo.