Cai Shi-rong, Chen Chuang-qi, Wang Zhao, He Yu-long, Cui Ji, Wu Wen-hui, Wu Hui, Zhan Wen-hua
Department of Gastrointestinal Surgery, Gastric Cancer Center, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China.
Zhonghua Yi Xue Za Zhi. 2008 Aug 26;88(33):2326-30.
To detect the expression of phosphatase of regenerating liver (PRL)-3 in primary gastric cancer tissues, evaluate its prognostic impact, and investigate the role of silencing PRL-3 expression by miRNA interference in gastric cancer growth.
Immunohistochemistry was used to measure the expression of PRL-3 in 137 gastric tumor samples. The overall survival rates of the patients with different PRL-3 expression levels were compared. Recombinant lentivirus expressing artificial PRL-3 miRNA, Lent. rPRL3-miRs, was established. Human gastric cancer cells of the line SGC7901 were cultured and transfected with Lent. rPRL3-miRs or blank vector, Lenti.rPRL-miR-neg, respectively. Un-transfected progenitor cells were used as controls. MTT assay was used to examine the proliferation of these cells. RT-PCR and Western blotting were used to detect the RNA and protein expression of PRL-3 in the SGC7901 cells. Thirty BALB/c mice were divided into 3 equal groups to be inoculated subcutaneously with SGC7901 cells transfected with Lent. rPRL3-miRs or blank vector, and control SGC7901 cells respectively. The growth of tumor was observed and the tumor sizes were measured 21 days later.
85 of the 137 gastric cancer samples (62%) showed high PRL-3 expression and 26 (19%) showed moderate and low PRL-3 expression. High PRL-3 expression was significantly correlated with tumor size (P < 0.01), infiltration depth (P < 0.01), lymph node metastasis (P < 0.01), hepatic metastasis (P < 0.01), adjacent organ invasion (P < 0.01), and TNM staging (P < 0.01). The median survival time of the patients with high PRL-3 expression in the primary tumor was 18.9 months, significantly shorter than those with moderate or low expression (39.1 and 74.3 months respectively, both P < 0.01). The growth rate of the SGC7901 cells transfected with the recombinant lentivirus expressing artificial PRL-3 miRNA was significantly lower than that of the blank vector-transfected group. The implanted tumor size of the Lenti. rPRL-3-miR-B transfection group was (1.92 +/- 0.18) cm(3), significantly smaller than those of the control and Lenti.rPRL-miR-neg groups [(4.86 +/- 0.38) and (4.74 +/- 0.39) cm(3) respectively, both P < 0.01].
High PRL-3 expression is associated with gastric cancer progression. Silencing of PRL-3 significantly suppresses the proliferation of gastric cancer cells and tumor growth. PRL-3 plays a key role in the growth of gastric cancer. PRL-3 should be considered as a potential therapeutic target.
检测再生肝磷酸酶（PRL）-3在原发性胃癌组织中的表达，评估其对预后的影响，并研究通过微小RNA干扰沉默PRL-3表达在胃癌生长中的作用。
采用免疫组织化学法检测137例胃肿瘤样本中PRL-3的表达。比较不同PRL-3表达水平患者的总生存率。构建表达人工PRL-3微小RNA的重组慢病毒Lent. rPRL3-miRs。培养人胃癌SGC7901细胞，分别用Lent. rPRL3-miRs或空载体Lenti.rPRL-miR-neg转染。未转染的祖细胞作为对照。采用MTT法检测这些细胞的增殖情况。采用RT-PCR和Western印迹法检测SGC7901细胞中PRL-3的RNA和蛋白表达。将30只BALB/c小鼠分成3组，每组数量相等，分别皮下接种经Lent. rPRL3-miRs或空载体转染的SGC7901细胞以及对照SGC7901细胞。观察肿瘤生长情况，并在21天后测量肿瘤大小。
137例胃癌样本中，85例（62%）PRL-3表达高，26例（19%）PRL-3表达为中度和低度。PRL-3高表达与肿瘤大小（P < 0.01）、浸润深度（P < 0.01）、淋巴结转移（P < 0.01）、肝转移（P < 0.01）、邻近器官侵犯（P < 0.01）及TNM分期（P < 0.01）显著相关。原发性肿瘤PRL-3高表达患者的中位生存时间为18.9个月，显著短于中度或低度表达患者（分别为39.1个月和74.3个月，P均< 0.01）。转染表达人工PRL-3微小RNA的重组慢病毒的SGC7901细胞生长速率显著低于空载体转染组。Lenti. rPRL-3-miR-B转染组植入肿瘤大小为（1.92±0.18）cm³，显著小于对照组和Lenti.rPRL-miR-neg组[分别为（4.86±0.38）cm³和（4.74±0.39）cm³，P均< 0.01]。
PRL-3高表达与胃癌进展相关。沉默PRL-3可显著抑制胃癌细胞增殖和肿瘤生长。PRL-3在胃癌生长中起关键作用。PRL-3应被视为潜在的治疗靶点。
