Relationship between HOX gene abnormality and investigation/metastasis of cancer cells in gynecologic malignancy

在本研究中，我们研究了HOX基因在卵巢和子宫内膜来源样本（正常卵巢和子宫内膜组织和癌细胞系）中的表达谱，这些癌细胞是否表达HOX基因，以及HOX基因的反义引入是否影响过表达HOX基因是否涉及癌细胞的侵袭/转移方式。首先，通过实时荧光定量PCR分析HOX基因在卵巢癌细胞（ES-2、SMOV-2、JHOC-6、SKOV-3、CAOV-3）和子宫内膜癌细胞（AN3CA、KLE、ISHIKAWA、HEC1A、HEC1B、RL-95）中的过表达情况。结果表明，与正常对照部分相比，部分HOX基因在肿瘤细胞中过度表达。特别是在卵巢癌和子宫内膜癌中均观察到HOXB13的过表达。其次，利用HOXB13作为靶基因反义导入癌细胞。构建反义导入载体后，用电镀法将其导入SKOV-3和AN3CA细胞。通过培养和筛选符合反义基因导入的细胞，将插入的细胞运送到基质室进行基质实验，研究反义DNA对癌细胞侵袭方式的影响。腔室中膜上细胞的培养和细胞计数显示，卵巢癌和子宫内膜癌的膜上癌细胞数量减少，而卵巢癌的长期培养显示癌细胞数量恢复。这些结果表明HOX基因表达在卵巢癌和子宫内膜癌中都起重要作用。

In this study, we investigated expression profile of the HOX genes in ovarian and endometrial derived samples (normal ovarian and endometrial tissues and cancer cell lines) whether these cancer cells express HOX genes and examined effect of antisense introduction of the HOX genes whether overexpressed HOX genes involve manner of invasion/metastasis of the cancer cells. First, overexpression of the HOX genes in ovarian cancer cells (ES-2, SMOV-2, JHOC-6, SKOV-3, CAOV-3) and endometrial cancer cells (AN3CA, KLE, ISHIKAWA, HEC1A, HEC1B, RL-95) were analyzed by real-time PCR assay. It demonstrated that some of HOX genes are overexpressed in the cancer cells compared with that of normal counter parts. Especially, overexpression of HOXB13 is observed in both ovarian and endometrial cancers. Secondary, HOXB13 was used as a target gene for antisense introduction to the cancer cells. After construction of a vector for antisense introduction, it was introduced into the SKOV-3 and AN3CA cells by electroplating method. Conforming of the introduction of the antisense gene by cultivation and selection, inserted cells were transported to the matrigel chamber for matrigel assay to investigate that antisense DNA influences manner of invasion of the cancer cells. Culture and cell count of the cells on the membrane in the chamber revealed decreased number of cancer cells on the membrane in both ovarian and endometrial cancers, however in ovarian cancer, long term cultivation showed recovery of the number of the cancer cells. These results indicate important role of the HOX gene expression in both ovarian and endometrial cancers.