MED12/HMGA2调控卵巢癌细胞衰老及耐药的机制研究

化疗耐药是导致卵巢癌治疗失败的主要原因。细胞衰老可分泌相关因子诱导周边肿瘤细胞耐药，其机制尚未阐明。我们前期研究发现，沉默MED12可下调衰老相关蛋白HMGA2的表达，诱导卵巢癌细胞发生衰老及耐药。据此我们提出假说：MED12通过调节HMGA2的表达调控卵巢癌细胞衰老并介导其化疗耐药。本研究拟采用免疫组化及ELISA技术检测卵巢癌标本中MED12的表达水平及腹水中衰老相关因子的表达，分析其与卵巢癌化疗效果及复发的关系；以CRISPR/Cas9或基因转染技术改变MED12表达，研究其对卵巢癌细胞体内外衰老和耐药的影响；通过基因芯片及ChIP-seq等方法，探索MED12调控的候选靶基因；以ChIP、双荧光素酶报告基因等技术，研究MED12调控HMGA2的分子机理。本研究将阐明MED12调控卵巢癌衰老和耐药的作用及其分子机理，为克服卵巢癌耐药、复发提供新策略。

Chemotherapy resistance is the main cause of failure in the treatment of ovarian cancer. It’s reported that senescent tumor cells could secrete related factors to induce drug resistance of peripheral tumor cells. The mechanism has not yet been clarified. Our previous studies have found that silencing MED12 could down-regulate the expression of senescence-related protein HMGA2 and induce cellular senecence and drug resistance of ovarian cancer cells. Therefore, we hypothesize that MED12 regulates cellular senescence of ovarian cancer cells by regulating the expression of HMGA2 and mediates their chemotherapeutic resistance. In this study, immunohistochemistry and ELISA will be used to detect the expression of MED12 in ovarian cancer specimens and the expression of senescence-related factors in ascites to investigate the clinical significance of MED12 in ovarian cancer. CRISPR/Cas9 and transfection will be used to study the function of MED12 in tumor cellular senescence and drug resistance of ovarian cancer in vitro and in vivo. Identification of downstream target genes of MED12 will be performed by microarray analysis and ChIP-seq assay. The mechanism of regulation of HMGA2 and downstream signaling by MED12 will be studied by ChIP, luciferase and other techniques. The study aims to investigate the effects of MED12 on tumor cellular senescence and drug resistance and its clinical significance in human ovarian cancer, to illustrate the function and mechanism of MED12 in regulating tumor cellular senescence and drug resistance, and to provide new strategies to overcome recurrence of ovarian cancer.