RNA干扰c-Met基因表达对非小细胞肺癌侵袭和迁移及顺铂敏感性的影响

[目的]研究c-Met基因干扰后对肺癌细胞株95D侵袭、迁移能力和化疗药物敏感性的影响。[方法]分别采用免疫组化SP法和WesternBlot技术检测肺癌组织及不同肺癌细胞株中Met蛋白的表达情况。将c-MetshRNA质粒转染人肺癌细胞株95D．通过WestemBlot检测转染效率;Transwell小室和划痕愈合实验测定细胞体外侵袭和迁移能力：四甲基偶氮唑法（Mar法）检测细胞的增殖情况及顺铂敏感性。[结果]Met蛋白在NSCLC组织中的表达明显高于癌旁正常组织,同时高侵袭性的肺癌细胞株（95D、801D）中Met蛋白的表达也较高。c-Met基因干扰能明显降低95D细胞的侵袭和迁移能力,并显著提高其对顺铂的敏感性。[结论]c-Met基因可作为一个重要的靶点应用于非小细胞肺癌治疗。     

miR-206过表达对甲状腺乳头状癌细胞增殖和迁移的影响

目的:明确微小RNA-206(microRNA-206,miR-206)对人甲状腺乳头状癌K1细胞增殖和迁移的影响,并初步探讨其可能的机制。方法:采用RT-qPCR技术检测各组细胞miR-206的表达水平;CCK-8实验和台盼蓝染色活细胞计数检测K1细胞的增殖能力;Transwell小室迁移实验检测K1细胞的迁移能力;利用生物信息学软件预测miR-206的靶基因;采用双萤光素酶报告基因检测法验证miR-206和c-Met的靶向关系;Western blot检测c-Met/AKT/mTOR通路中各蛋白水平的变化。结果:K1细胞瞬时转染miR-206 mimic后,RT-qPCR检测实验组K1细胞中miR-206的相对表达量较空白组和阴性对照组明显升高(P<0.01);CCK-8及台盼蓝拒染实验表明,转染miR-206mimic的实验组中K1细胞的增殖较空白组和阴性对照组明显受到抑制(均P<0.01);Transwell实验发现,实验组K1细胞的迁移数低于空白组和阴性对照组(P<0.01);双萤光素酶报告基因检测表明,miR-206可负调控靶基因c-Met的表达;West...     

非小细胞肺癌放疗抵抗研究进展

放射治疗是非小细胞肺癌的重要治疗手段之一，而肿瘤放疗抵抗限制了放疗的疗效。提高放疗疗效仍是目前非小细胞肺癌患者治疗中亟待解决的重要难题之一。研究PI3K/AKT/mTOR信号通路、肿瘤干细胞、微小RNA、HGF/c-Met、常规分割放疗等在非小细胞肺癌放疗抵抗中的作用机制，对提高非小细胞肺癌患者的生存率及改善非小细胞肺癌患者的预后具有重要意义。     

Targeting slug-mediated non-canonical activation of c-Met to overcome chemo-resistance in metastatic ovarian cancer cells

Metastasis-associated drug resistance accounts for high mortality in ovarian cancer and remains to be a major barrier for effective treatment. In this study, SKOV3/T4, a metastatic subpopulation of ovarian cancer SKOV3 cells, was enriched to explore potential interventions against metastatic-associated drug resistance. Quantitative genomic and functional analyses were performed and found that slug was significantly increased in the SKOV3/T4 subpopulation and contributed to the high resistance of SKOV3/T4. Further studies showed that slug activated c-Met in a ligand-independent manner due to elevated levels of fibronectin and provoked integrin α V function, which was confirmed by the significant correlation of slug and p-Met levels in 121 ovarian cancer patient samples. Intriguingly, c-Met inhibitor(s) exhibited greatly enhanced anti-cancer effects in slug-positive ovarian cancer models both in vitro and in vivo. Additionally, IHC analyses revealed that slug levels were highly correlated with reduced survival of ovarian cancer patients. Taken together, this study not only uncovers the critical roles of slug in drug resistance in ovarian cancer but also highlights a promising therapeutic strategy by targeting the noncanonical activation of c-Met in slug-positive ovarian cancer patients with poor prognosis.     

肝细胞生长因子及其受体c-Met在子宫内膜癌中的表达

目的　探讨肝细胞生长因子 (HGF)及其受体c Met蛋白表达与子宫内膜癌临床病理特征的关系。方法　应用免疫组化法检测 2 0例正常子宫内膜 ,4 6例子宫内膜癌组织中HGF及c Met蛋白的表达。结果　①正常子宫内膜HGF及c Met在增生期内膜的表达明显高于分泌期子宫内膜的表达。②子宫内膜癌c Met表达与临床分期、病理分级明显相关 (P <0 .0 5 ) ,Ⅰ、Ⅱ期低于Ⅲ、Ⅳ期 ,G1、G2 级低于G3 级。与年龄、肌层浸润、淋巴结转移无关 (P>0 .0 5 )。半定量分析 :HGF表达与子宫内膜癌Ⅲ +Ⅳ期有关 (P <0 .0 5 )。结论　HGF/c Met系统在子宫内膜癌的侵袭进展中起重要作用 ,c Met可能成为判断子宫内膜癌预后的良好参考指标。HGF及c Met联合检测对于判断子宫内膜癌预后有重要意义     

人晶状体上皮细胞HGF与c-Met的表达对增殖和凋亡的影响

目的：探讨肝细胞生长因子(hepatocyte growth factor,HGF)在人晶状体上皮细胞(lens epithelial cell,LEC)的表达及其对晶状体上皮细胞的促增殖和抑制凋亡的作用。方法：取原代培养人晶状体上皮细胞,应用RT-PCR,Western blot检测HGF,C-Met的mRNA及蛋白的表达,应用MTT法检测HGF对晶状体上皮细胞增殖的影响,Western-blot检测：Bcl-2的蛋白表达,以揭示其对晶状体上皮细胞凋亡的影响。结果：HGF和c-Met在人晶状体上皮细胞有表达,HGF有促进晶状体上皮细胞增殖的作用,并能抑制凋亡产生。结论：HGF参与人晶状体上皮细胞的增殖代谢过程,与晶状体上皮细胞的增殖相关。     

c-Met在肺癌中的研究进展

c-Met是酪氨酸激酶受体的一种,由MET基因编码产生.肝细胞生长因子(HGF)作为c-Met唯一的天然配体,与其结合后激活相关下游通路如PI3K、MAPK和STAT 3等,参与肿瘤的增殖、迁移、侵袭等方面.c-Met主要激活形式包括有配体依赖的自分泌或旁分泌机制和非配体依赖机制等,许多恶性肿瘤包括肺癌在内均存在c-M...     

色素上皮衍生因子通过抑制PI3K/Akt通路降低宫颈癌HeLa细胞活性和侵袭相关蛋白表达

目的：探讨色素上皮衍生因子（PEDF）通过抑制磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（Akt）信号通路,从而下调人宫颈癌HeLa细胞的活性和侵袭相关蛋白c-Met表达的相关机制。方法：选取本院病理已确诊的40例宫颈癌样本及对应的40例癌旁正常组织样本,通过免疫组织化学法检测PEDF、PI3K和p-Akt蛋白的表达。在细胞水平分为正常宫颈上皮细胞HCerEpiC组、HeLa组、HeLa+PEDF组、HeLa+PEDF+PI3K/Akt激活剂组和HeLa+PI3K/Akt抑制剂组。通过蛋白质印迹（Western blotting）检测各组细胞内PEDF、PI3K、p-Akt、Akt及c-Met蛋白的表达水平,CCK-8法检测各组细胞的活性。结果：宫颈癌样本和HeLa细胞中的PI3K/Akt通路显著激活,且PEDF表达显著下降（P＜0.01）。PEDF可以显著抑制HeLa细胞中PI3K/Akt激活和细胞活性以及侵袭相关蛋白水平的增加（P＜0.01）。Akt激活剂可以逆转PEDF对HeLa细胞活性和侵袭蛋白表达的影响,而激活PI3K无法改变PEDF的作用。单纯的药物抑制PI3K/Akt通路不足以模拟PEDF对HeLa细胞的作用。结论：PEDF通过PI3K/Akt双重抑制作用降低HeLa细胞活性和侵袭相关蛋白表达,这为临床上PEDF治疗宫颈癌提供了新的思路和理论依据。     

Prognostic Factors and Recurrence of Small Hepatocellular Carcinoma after Hepatic Resection or Microwave Ablation: A Retrospective Study

Purpose This study aims to analyze the long-term therapeutic results of small HCC less than 5 cm in diameter after microwave ablation (MA) or hepatic resection (HR) and choose factors that could predict metastasis and recurrence of small HCC. Materials and Methods The metastasis and recurrence of 194 patients with one HCC less than 5 cm in diameter who underwent curative HR or MA between January 1995 and December 2004 were reviewed retrospectively; immunohistochemistry was used to analyze the expressions of VEGF, bFGF, and c-Met in HCC tissues. Posttreatment prognostic factors were evaluated by multivariate analysis using Cox’s proportional hazards model. The variables included the expressions of these three proteins in HCC tissues, the clinical and pathologic characteristics of the patients. Results The retrospective study showed that 1-, 3-, and 5-year disease-free survival rates of patients with single HCC of diameter <5 cm were 71.3, 57.0 and 32.5%, respectively. Furthermore, 1-, 3-, and 5-year disease-free survival rates of the patients in MA group and resection group were 72.8, 54.0 and 33.0%; 68.5, 60.0, and 25.6%, respectively. There was no significant difference in disease-free survival rates between these two groups. The result of multivariate analysis showed that differentiation degree of HCC and the expressions of VEGF and c-Met in HCC tissues could be as the independent prognostic factors affecting metastasis and recurrence in patients with small HCC, whereas the methods of therapy had no impact on prognosis. Conclusions The metastasis and recurrence rate after MA is similar to that after HR, and the methods of therapy do not affect the prognosis of small HCC. The metastasis and recurrence of patients with small HCC will differ depending on tumor differentiation, expressions of VEGF and c-Met in HCC tissues.