ARPP-19调控细胞周期促进肝癌细胞增殖及中药肝积方的干预

Hepatocellular carcinoma(HCC) still belongs to the most common malignancy affecting human health seriously, which challenges the development of efficient diagnosis and treatment measures urgently. One important therapeutic target is to inhibit HCC cell proliferation. Recent studies demonstrate that cAMP regulated phosphoprotein 19 (ARPP-19) play a pivotal role for cell mitosis G2/M transition. And overexpression of ARPP-19 in HCC was detected in our previous small sample experiment. Traditional Chinese medicine Ganji recipe has been used in clinic for a long term in Tumor Department of Longhua Hospital, with function of improving survival time of HCC patients, while the unclear underlying mechanisn limits the extending application..Therefore, this study will compare the expression and activity of ARPP-19 in human HCC and the adjacent tissues, to analyze the correlation with HCC. Then in vitro and in vivo experiments will be performed to investigate ARPP-19 function in HCC cell cycle control, proliferation regulation and tumorigenisis, as well as the signal pathway. In addition, Ganji recipe will be applied to in vivo HCC model to observe the intervening impact, and more significant, to explore the HCC-suppressing mechanism of Ganji recipe from the point of mitosis cell cycle regulation and proliferation..This study would focus on new insight into the pathogenesis of HCC and provide new molecule target for diagnosis and therapy, as well as reveal the therapeutic biological mechanism of the Ganji recipe for HCC, which may help develop and spread the application of traditional Chinese medicine in HCC comprehensive therapy.

肝细胞性肝癌（HCC）严重危害人类健康，目前诊疗措施仍不足以控制该疾病。抑制HCC细胞分裂增殖是重要治疗靶向。在前期小样本试验中发现环腺苷酸调节磷酸化蛋白-19（ARPP-19）在HCC组织表达升高，而近期报道该分子是进入分裂中期的关键调控分子。本单位据肝癌病机"脾虚为本，气滞血瘀、湿热、毒邪为标"组成中药肝积方，长期应用于临床可抑制肿瘤生长，有效延长肝癌患者生存期，但机制不明限制其推广应用。.基于此，本项目首先应用临床组织样本研究ARPP-19表达活化与HCC相关性；进而通过体外、体内HCC模型研究ARPP-19对HCC细胞周期、增殖、成瘤的影响及相关途径；同时基于ARPP-19调控细胞周期和增殖方面探讨肝积方治疗HCC的药效机制。.通过本项目有望揭示HCC发生发展新机制，增加HCC诊疗新靶标，为中医HCC辩证治则理论提供现代生物学释义，为肝积方治疗HCC的推广提供科学依据。

目前仍然缺乏肝细胞性肝癌（HCC）有效治疗方法，中药肝积方在本单位肿瘤科长期应用于临床，可有效提高肝癌病人生存期，但其机制有待探讨。研究表明环腺苷酸调节的磷酸化蛋白-19（ARPP-19）是细胞进入分裂期的重要调控分子，在胚胎期及一些恶性细胞中表达较多。.. 本项目首先检测36对HCC、癌旁肝组织中ARPP-19表达水平，发现ARPP-19在人HCC组织表达上调，并与肿瘤大小相关。应用HCC细胞体外试验表明，抑制该分子表达导致细胞增殖下降，克隆形成能力降低，细胞在G2/M期被阻滞增加，而与M期相关的分子CDC2等表达有所变化，增加其表达促进细胞增殖。下调ARPP-19抑制了细胞分裂相关底物、周期依赖性激酶CDC2的活化。证明ARPP-19可调控M期分裂从而影响了HCC细胞增殖。裸鼠皮下种植过表达ARPP-19的HCC细胞，较对照细胞成瘤体积显著增加，Ki67在瘤组织中的阳性率也明显增加，表明ARPP-19表达水平与肝癌细胞成瘤性、肿瘤增殖生长调控相关。而中药肝积方干预可抑制移植瘤生长，使HCC肿瘤组织中ARPP-19表达及Ki67阳性率下降，提示下调ARPP-19表达是其作用机制之一。.. 本研究结果表明ARPP-19可作为HCC诊断、治疗选择和判断预后的辅助指标，进一步深入研究有望进行相关新药开发。药效机制不清严重影响中医药临床推广应用，通过本项目明确中药复方肝积方可抑制HCC发生发展，其作用途径之一为抑制促分裂增殖分子ARPP-19及其信号通路，为中医药治疗HCC提供依据。