SLC12A1基因调控肝癌循环肿瘤细胞异质性及其在肝癌转移复发中的相关机制研究

The circulating tumor cells, as the core part of matastasis did arouse much concern of oncologist. As a vital part of the CTC researchs, heterogeneity of CTC and functions of CTC subsets have been an immediate areas of research focus, though full of frustrations.In our previous study, a system called “HCC-CTC 3D-hanging drop culture method” was devised, and CTCs clones shows distinct phenotypic states in 3D-gel (we call them MC or OC in short). Next, the gene profiling of the two types was revelled by single-cell sequencing, indicating SLC12A1 is the markedly upregulated in MC type which showed moer powerful proliferative capability. Further study clarified that SLC12A1 could pormoted liver cancer cell proliferation. Based on those issues, we proposed a reseach plan on SLC12A1 regulating role of circulating tumor cells heterogeneity and its mechanism for hepatocellular carcinoma metastasis. For one thing, the relationship between MC or OC counting number and prognosis would be illustrated by using “HCC-CTC 3D-hanging drop culture method”. For another, we would revel the phenotype alternation of CTC clones after intervening SLC12A1 expression. The study will demonstrates a link betweencell number of different CTC subsets and cancer metastasis. Also it will provide experimental bases for clinical detection and cancer prognosis.

作为肝癌转移的核心环节，循环肿瘤细胞引起了肿瘤学界的广泛关注。目前CTC异质性及其亚群功能的探索是研究热点，也是难点。在前期研究中我们构建了一种“HCC-CTC 3D悬滴法培养体系”，这一体系培养出的肝癌CTC呈现出两种表型完全不同的克隆形态(我们将其命名为MC型克隆与OC型克隆)。对两种克隆进行单细胞测序，发现在增殖能力更强的MC型克隆中SLC12A1是上调最为显著的分子，进一步实验证实SLC12A1可以促进肝癌细胞增殖。基于上述结果我们提出研究SLC12A1基因调控肝癌循环肿瘤细胞异质性这一课题，拟通过现有的CTC培养体系阐明MC与OC克隆数与肝癌患者预后之间的关系，并通过干预SLC12A1的表达研究该基因对CTC生物学表型的影响。该研究将有助于阐明不同亚群CTC数目与肿瘤转移之间的关联，为肝癌临床检测与预后判断提供实验依据。

肝癌严重威胁着人类健康，其主要致死原因是肿瘤转移。作为肿瘤转移的核心环节，循环肿瘤细胞在引起了肿瘤学界的广泛关注。而CTC异质性及其亚群功能的研究是其中的一个研究热点，也是难点。在前期研究中我们构建了一种“HCC-CTC 3D悬滴法培养体系”，这一体系培养出的肝癌CTC呈现出两种表型完全不同的克隆形态(我们将其命名为MC型克隆与OC型克隆)。对两种克隆进行单细胞测序，发现在增殖能力更强的MC型克隆中SLC12A1是上调最为显著的分子，进一步实验证实SLC12A1可以促进肝癌细胞增殖。基于上述结果我们提出研究SLC12A1基因调控肝癌循环肿瘤细胞异质性这一课题，拟通过现有的CTC培养体系阐明MC与OC克隆数与肝癌患者预后之间的关系，并通过干预SLC12A1的表达研究该基因对CTC生物学表型的影响。该研究将有助于阐明不同亚群CTC数目与肿瘤转移之间的关联，为肝癌临床检测与预后判断提供实验依据。