Abstract：
Despite the knowledge about numerous genetic mutations essential for the progression of low-grade serous ovarian carcinoma (LGSOC), the specific combination of mutations required remains unclear. Here, we aimed to recognize the oncogenic mutations responsible for the stepwise development of LGSOC using immortalized HOVs-cyst-1 cells, developed from ovarian serous cystadenoma cells, and immortalized via cyclin D1, CDK4R24C, and hTERT gene transfection. Furthermore, oncogenic mutations, KRAS and PIK3CA, were individually and simultaneously introduced in immortalized HOV-cyst-1 cells. Cell functions were subsequently analyzed via in vitro assays. KRAS or PIK3CA double mutant HOV-cyst-1 cells exhibited higher cell proliferation and migration capacity than the wild-type cells, or those with either a KRAS or a PIK3CA mutation, indicating that these mutations play a causative role in LGSOC tumorigenesis. Moreover, KRAS and PIK3CA double mutants gained tumorigenic potential in nude mice, whereas the cells with a single mutant exhibited no signs of tumorigenicity. Furthermore, the transformation of HOV-cyst-1 cells with KRAS and PIK3CA mutants resulted in the development of tumors that were grossly and histologically similar to human LGSOCs. These findings suggest that simultaneous activation of the KRAS/ERK and PIK3CA/AKT signaling pathways is essential for LGSOC development.
摘要：
尽管了解了许多对低级别浆液性卵巢癌（LGSOC）进展至关重要的基因突变，所需突变的具体组合仍不清楚.这里,我们的目的是使用永生化的HOVs-cyran-1细胞识别负责LGSOC逐步发展的致癌突变，从卵巢浆液性囊腺瘤细胞发展而来的，并通过细胞周期蛋白D1、CDK4R24C永生化，和hTERT基因转染。此外，致癌突变，KRAS和PIK3CA,分别同时引入永生化HOV-囊肿-1细胞。随后通过体外测定分析细胞功能。KRAS或PIK3CA双突变HOV-囊肿-1细胞比野生型细胞表现出更高的细胞增殖和迁移能力，或者有KRAS或PIK3CA突变的人，表明这些突变在LGSOC肿瘤发生中起致病作用。此外，KRAS和PIK3CA双突变体在裸鼠中获得了致瘤潜力，而具有单个突变体的细胞没有表现出致瘤性的迹象。此外，用KRAS和PIK3CA突变体转化HOV-囊肿-1细胞导致肿瘤的发展，在组织学上与人类LGSOCs大致相似。这些发现表明，KRAS/ERK和PIK3CA/AKT信号通路的同时激活对于LGSOC的发展至关重要。
