本研究将进行卵巢癌肿瘤内异质性的遗传生物学特点分析及相关分子靶标的筛选。流式分选获得遗传背景相同的卵巢癌细胞系中存在的侧群细胞、CD117+细胞;有限稀释法对卵巢癌细胞进行单细胞培养，分离获得异质性亚群。对分选获得的3种异质性细胞体外培养扩增，建立相关的动物模型。比较它们在细胞形态、分化能力、细胞周期、增殖能力、抗药性及裸鼠异体移植成瘤能力等方面的差异；利用外显子和/或全基因组测序，比较基因组杂交研究基因序列和基因拷贝数目的变化，并进行染色体畸变分析；通过蛋白芯片，甲基化芯片，miRNA芯片研究与卵巢癌肿瘤内异质性相关的分子及调控机制。最后应用卵巢癌原发/转移/复发病灶的组织芯片以及来自同一患者的原发肿瘤细胞系和复发肿瘤细胞系，验证筛选出的分子标记物是否在卵巢癌转移，复发和耐药过程中产生差异性，并明确其临床意义，为提高卵巢癌的诊治水平提供新的诊治思路。

Our research will focus on the genetic and biological characteristices analysis and potential molecular biomarkers screening of intratumor heterogeneity in epithelial ovarian cancer.SP cells and CD117+ cells will be obtained by flow cytometry;subclonal cancer cells will be obtained by single cell culture via limited dilution method. Three pairs of cell lines will be cultured in vitro and used to establish animal model, to compare their differences in morphology,proliferation,differentiation, drug resistance,and tumor formation in vivo.Whole genome sequencing and comparative genomic hybridization will be used to analyze gene mutations, gene copy number variations, and chromosome aberration. Protein chip, methylation chip,and miRNA chip will be adopted to screen the potential molecular biomarkers invovled in intratumor heterogeneity. Finally, tissue microarrys and cancer cell lines primarily cultured from ovarian cancer patients will be used to verify the functions of screened molecular biomarkers invovled in metastasis and recurrence of ovarian cancer.