抗癌新化合物盐霉素通过调控ATP2A3抗肿瘤干细胞的机理

In recent years, researcher found that salinomycin can highly kill a variety of cancer stem cells (CSCs), but the mechanism underlying anti-CSC remains clear. In our preliminary study, we found that salinomycin could increase the expression of ATP2A3 gene that caused endoplasmic reticulum stress and its downstream effects（cause CSC apoptosis, restrain the proliferation and differentiation of CSC, etc）, which may be one of leading anti-CSC mechanisms. The specific aims are to detect the intracellular Ca2+ concentration changes of prostate CSCs before and after salinomycin treatment. Meanwhile, the expression of ATP2A3 gene will be quantified by RT-PCR to further confirm the relationship between ATP2A3 gene expression and Ca2+ concentration in CSC.Also, we will investigate the relationship between ATP2A3 gene expression and OCT4, SOX2, NANOG genes expression,and confirm the highly expression of the ATP2A3 gene could cause CSC apoptosis in vitro and in vivo . Furthermore, we will elucidate the mechanism of salinomycin anti-cancer wherein the CSC proliferation, differentiation and infiltration may be influenced by these OCT4, SOX2 and NANOG genes through in vitro CSC culture model and NOD/SCID model in combination with the siRNA technology. Thereby clarify the mechanism of salinomycin anti-CSC through upregulation of ATP2A3, to provide a theoretical basis that salinomycin developed into a new drug of anti-Cancer stem cells.

近年来研究者发现盐霉素能高效杀死多种肿瘤干细胞(CSC)，但其作用机制仍不明确。申请者前期研究首次发现盐霉素能上调ATP2A3基因表达并引起内质网应激及其下游效应（诱导CSC凋亡，抑制更新、分化等）可能是其抗CSC作用机制。本课题拟采用激光共聚焦检测盐霉素作用前后CSC内Ca2+浓度的变化，RT-PCR检测ATP2A3基因表达与CSC内Ca2+浓度变化的关系，运用基因转染技术明确ATP2A3与OCT-4、SOX2、NANOG三个基因表达的关系，并证实体内外CSC中ATP2A3基因的高表达均能诱导CSC凋亡；采用siRNA技术下调体外CSC及NOD/SCID小鼠体内CSC中OCT-4、SOX2、NANOG基因的表达，明确上述三个基因的下调能在体内外抑制CSC的增值、分化及浸润；从而阐明盐霉素通过上调ATP2A3基因抗CSC的作用机制，为把盐霉素开发成抗肿瘤干细胞新药提供理论依据。

近年来研究者发现盐霉素能高效杀死多种肿瘤干细胞(CSC)，但其作用机制仍不明确。申请者前期研究首次发现盐霉素能上调ATP2A3 基因表达并引起内质网应激及其下游效应（诱导CSC 凋亡，抑制更新、分化等）可能是其抗CSC 作用机制。本课题拟采用激光共聚焦检测盐霉素作用前后CSC 内Ca2+浓度的变化，RT-PCR 检测ATP2A3 基因表达与CSC 内Ca2+浓度变化的关系，运用基因转染技术明确ATP2A3 与OCT-4、SOX2、NANOG 三个基因表达的关系，并证实体内外CSC 中ATP2A3 基因的高表达均能诱导CSC 凋亡；采用siRNA 技术下调体外CSC 及NOD/SCID 小鼠体内CSC 中OCT-4、SOX2、NANOG 基因的表达，明确上述三个基因的下调能在体内外抑制CSC 的增值、分化及浸润；从而阐明盐霉素通过上调ATP2A3 基因抗CSC 的作用机制，为把盐霉素开发成抗肿瘤干细胞新药提供理论依据。