人脂肪来源间充质干细胞诱导人乳腺癌MCF-7细胞上皮间质转化

目的探讨人脂肪来源间充质干细胞(hAD-MSCs)在乳腺肿瘤微环境中对乳腺癌细胞的影响。方法采用Transwell体系间接共培养人乳腺癌细胞系(MCF-7)和hAD-MSCs,对共培养后的肿瘤细胞进行细胞形态,EMT相关标志以及肿瘤特性的检测。结果与对照组相比,共培养之后的MCF-7细胞发生上皮间质转化。与hAD-MSCs共培养能明显促进乳腺癌细胞的侵袭和迁移,但对细胞周期和增殖能力没有明确的影响。结论人脂肪来源间充质干细胞可诱导乳腺癌细胞上皮间质转化。     

15-Hydroxyprostaglandin dehydrogenase associates with poor prognosis in breast cancer, induces epithelial-mesenchymal transition, and promotes cell migration in cultured breast cancer cells

Breast cancer is the most frequent cancer and the leading cause of cancer-related deaths in women worldwide. The prognosis of breast cancer is tightly correlated with the degree of spread beyond the primary tumour. Arachidonic acid (AA) and prostaglandin E(2) (PGE(2)) are known to regulate tumour metastasis enabling epithelial-mesenchymal transition (EMT). However, the detailed role of 15-hydroxyprostaglandin dehydrogenase (HPGD), the key enzyme degrading prostaglandin E(2) , remains unclear in breast cancer. Here, we show that HPGD mRNA is overexpressed in a subset of clinical breast cancers compared to normal breast tissue samples and that high HPGD mRNA expression associates with poor prognosis. Immunohistochemical staining of primary breast cancer and lymph node metastasis tissue samples confirmed high HPGD protein expression in 20% of the samples, as well as associated HPGD expression with aggressive characteristics, such as increased risk of disease relapse and shorter disease-free survival. Results from cultured cells indicated abundant HPGD expression in highly metastatic breast cancer cells, and impairment of HPGD expression using RNA interference led to a significant decrease in transforming growth factor-β signalling, in cellular arachidonic acid levels as well as in cell migration. Furthermore, gene expression microarray analysis followed by quantitative RT-PCR validation showed that HPGD silencing decreased aryl hydrocarbon receptor signalling and induced mesenchymal-epithelial transition. In conclusion, our results indicate that HPGD is highly expressed in metastatic and aggressive breast cancer and promotes EMT and migration in breast cancer cells.     

肝细胞生长因子促进HeLa细胞上皮间质转化

目的探讨肝细胞生长因子对宫颈癌He La细胞上皮间质转化的影响。方法以不同浓度的HGF(0、5、10、20和40 ng/m L)处理细胞24 h或20 ng/m L肝细胞生长因子(HGF)处理细胞不同时间(0、6、12、24和48 h),MTT检测细胞活力;用HGF(20 ng/m L)处理细胞24 h,倒置显微镜观察He La细胞形态,划痕实验检测细胞的迁移及Transwell实验检测细胞的侵袭能力。Western blot检测上皮细胞的标志蛋白E-Cadherin及转录因子snail和间质细胞的表面标志蛋白vimentin的表达,HGF/c-Met信号通路分子的表达情况。结果 HGF促进He La细胞增殖,其中以20 ng/m L或20 ng/m L处理24 h生存率达到最高峰;此外,HGF可以促进He La细胞迁移、侵袭及上皮间质转化,并促进c-Met磷酸化,并且激活p-ERK、p-AKT以及促进snail的蛋白表达(P0.05或P0.01)。结论 HGF可以促进He La细胞上皮间质转化,其机制可能是通过HGF/c-Met/MAPK或HGF/c-Met/AKT通路上调snail表达,进而调控E-cadherin或vimentin表达有关。     

IMP3 expression is associated with epithelial-mesenchymal transition in breast cancer

IMP3 plays an important role in tumor invasion and metastasis, to which epithelial to mesenchymal transition (EMT) also contributes. The purpose of this study was to investigate whether IMP3 can regulate invasion and metastasis through EMT in breast cancers. The protein expression levels of IMP3 and EMT markers were analyzed by immunohistochemistry in 180 paraffin-embedded human breast tissue samples. There was an inverse correlation of IMP3 with E-cadherin protein expression (P = 0.042). IMP3 expression directly correlated with both Slug (P = 0.004) and vimentin (P < 0.001). Changes in E-cadherin, vimentin, and Slug mRNA and protein levels were examined by quantitative real-time reverse polymerase chain reaction (qRT-PCR) and western blotting. Overexpression of IMP3 reduced the expression of E-cadherin and upregulated Slug and vimentin in transfected cells. In contrast, knocking down IMP3 had the opposite expression of the three proteins. Ribo-immunoprecipitation qPCR revealed that IMP3 binds Slug mRNA directly. In a transwell assay, overexpression of Slug rescued the cell migration and invasion caused by silencing IMP3 in MDA-MB-231 cells. On the other hand, knockdown of Slug in T47D-IMP3 cells could also have the opposite change. Our results strengthen the association of IMP3 with the regulation of EMT. Slug is a functional target of IMP3. IMP3 could therefore promote invasion and migration through the EMT in breast cancer cells.     

MiR-223 inhibited cell metastasis of human cervical cancer by modulating epithelial-mesenchymal transition

Accumulating evidence have emerged important roles for microRNAs (miRNAs) participating in epithelial-mesenchymal transition (EMT) process and are associated with metastasis in cervical cancer. We hypothesized that miR-223 played an important role in cell metastasis of cervical cancer. Here, we found miR-223 was downregulated in human cervical cancer cell lines and clinical tumor tissues. Result of wound healing and cell migration assays revealed that miR-223 inhibited cell migration, whereas miR-223-in showed the opposite effect. In terms of mechanism, miR-223 influenced the expression of the EMT-associated proteins by upregulating the epithelial markers E-cadherin and α-cadherin and downregulating the mesenchymal marker vimentin. In conclusion, miR-223 inhibited cell metastasis of human cervical cancer by modulating epithelial-mesenchymal transition.     

Increased expression of hepatoma-derived growth factor correlates with poor prognosis in human nasopharyngeal carcinoma

Wang S & Fang W
(2011) Histopathology 58 , 217–224
Increased expression of hepatoma‐derived growth factor correlates with poor prognosis in human nasopharyngeal carcinoma Aims: To examine the correlation between hepatoma‐derived growth factor (HDGF) expression and clinicopathological data in nasopharyngeal carcinoma (NPC), including patient survival. Methods and results: Using real‐time polymerase chain reaction (PCR) and Western blot, mRNA and protein expression of HDGF was detected in normal nasopharyngeal tissues, NPC tissues and cell lines. HDGF levels were determined further by an immunohistochemical analysis in a retrospective series consisting of 160 primary NPC tissues and 71 non‐cancerous nasopharynx tissues. Overexpressed mRNA and HDGF protein was present in NPC. By immunohistochemical analysis, we found that 53.8% (86 of 160) and 19.4% (32 of 160) of NPC biopsy specimens showed higher HDGF expression of the nucleus and cytoplasm, respectively. Statistical analysis showed that the higher expression of nuclear HDGF was associated significantly with T stage (P = 0.005) and clinical stage (P = 0.038), but there was no association with lymph node (P = 0.059) or distant metastasis (P = 0.563). Patients with increased HDGF expression levels had poorer overall survival rates than those with low expression of HDGF levels (P = 0.006). Multivariate analysis revealed that high expression of nuclear HDGF was an independent prognostic indicator of patient survival. Conclusions: Increased nuclear expression of HDGF is a potential unfavourable prognostic factor for patients with NPC.     

表皮生长因子促进电压门控钠通道Nav1.5表达及人乳腺癌细胞的侵袭

目的 探讨表皮生长因子(EGF)是否调节乳腺癌MDA-MB-231细胞电压门控钠通道(VGSC)Nav1.5亚型基因的表达及其可能的信号分子传导途径.方法 用Matrigel侵袭、免疫荧光定位、实时荧光定量RT.PCR(RFQ-PCR)和Western blot等方法,分别检测或探讨EGFR和Nav1.5蛋白在细胞中的表达和定位、EGF和Nav1.5对细胞侵袭的作用、EGF对Nav1.5 mRNA和蛋白水平的影响以及P13K在EGF增加侵袭中的作用.结果 MDA-MB-231细胞高表达EGF受体和Nav1.5蛋白,EGF增加细胞的侵袭达51.0%±2.6%,VGSC抑制剂Tetrodotoxin(TTX)10 μmol/L阻滞EGF诱导的细胞侵袭(P＜0.05);EGF增加Nav1.5 mRNA水平为(128±4)倍(P＜0.05),Nav1.5蛋白水平有39%±4%上升(P＜0.05).P13K抑制剂Wortmannin抑制EGF诱导的Nav1.5 mRNA和蛋白水平的增加,亦抑制EGF诱导的细胞侵袭.结论 EGF上调乳腺癌MDA-MB-231细胞VGSC Nav1.5亚型mRNA和蛋白的表达,促使乳腺癌细胞的侵袭,P13K参与EGF诱导的Nav1.5的表达和细胞的侵袭.     

转录因子Runx2对人乳腺癌MDA-MB-231细胞上皮-间充质转化能力的影响

目的:建立稳定敲减Runt相关转录因子2(Runx2)表达的人乳腺癌MDA-MB-231细胞系,探讨Runx2对MDA-MB-231细胞上皮-间充质转化(EMT)、转移和侵袭能力的影响。方法:利用LV-Runx2-RNAi慢病毒载体感染MDA-MB-231细胞,构建稳定低表达Runx2的MDA-MB-231细胞株,检测比较Runx2表达水平对MDA-MB-231细胞的形态及E-cadherin、N-cadherin、β-catenin和基质金属蛋白酶9(MMP-9)蛋白表达的影响。侵袭实验和软琼脂集落形成实验分析比较抑制Runx2表达对乳腺癌细胞侵袭和非锚定生长能力的效应。结果:E-cadherin蛋白表达在敲减Runx2表达的MDA-MB-231细胞中明显高于常规MDA-MB-231细胞(P0.05),而N-cadherin、β-catenin和MMP-9蛋白表达在敲减Runx2表达的MDA-MB-231细胞中明显低于常规MDA-MB-231细胞(P0.05)。敲减Runx2表达的MDA-MB-231细胞侵袭和非锚定生长能力明显低于常规MDA-MB-231细胞(P0.05)。结论:在人乳腺癌MDA-MB-231细胞中抑制Runx2表达可以通过调控EMT相关蛋白的表达进而抑制乳腺癌细胞的EMT过程,从而对细胞的侵袭和转移起负调控作用。     

成纤维生长因子受体信号通路与横纹肌肉瘤上皮间叶转化

横纹肌肉瘤是起源于骨骼肌的间叶源性肿瘤,常发生于15岁以下的儿童和青少年。由于不同类型横纹肌肉瘤的组织学特征、发病年龄、部位和预后各具差异,迄今为止横纹肌肉瘤的分子机制仍不明确。近年来,关于其分子机制的研究主要集中于关键基因和信号通路等方面,研究发现成纤维生长因子受体信号通路与横纹肌肉瘤的生长及转移密切相关,在多数上皮源性肿瘤中已证实上皮间叶转化促进了肿瘤增生与转移,因此笔者提出在横纹肌肉瘤中上皮间叶转化可能通过成纤维生长因子受体信号通路促进肿瘤增生和转移的假设,结合成纤维生长因子受体信号通路和上皮间叶转化来试着解释横纹肌肉瘤的发生、发展,以期为其治疗和预后评估提供一定的帮助。     

血管内皮生长因子C在人乳腺癌细胞株MDA-MB-231中的表达

目的：研究血管内皮生长因子C(VEGF-C)在人乳腺癌细胞中的体外表达情况，探讨其在肿瘤发生、发展中的作用。方法：运用半定量RT—PCR和免疫组织化学的方法分别检测了VEGF-CmRNA和VEGF-C在人乳腺癌细胞株MDA—MB-231中的表达。结果：半定量RT-CR检测到VEGF-CmRNA以较高水平表达，免疫组化结果显示MDA—MB-231细胞的胞浆中有棕黄色阳性颗粒，而阴性对照细胞的胞浆中则均无阳性颗粒。结论：人乳腺癌细胞株MDA-MB-231细胞在体外能够转录VEGF—CmRNA并翻译合成相应的蛋白。     

Role of vascular endothelial growth factor during breast cancer

We review the results of experimental and clinical observations on neoangiogenesis in patients with breast cancer. Vascular endothelial growth factor is an important positive regulator of this process. Experiments showed the possibility of using various direct and indirect antiangiogenic means in the therapy of breast cancer, but clinical efficiency of these methods was not proved. Expression of vascular endothelial growth factor can serve as a prognostic criterion in breast cancer. Antiangiogenic preparations should not be used as monotherapy, but as the treatment complementary to standard therapy.     

Placenta growth factor promotes migration through regulating epithelial-mesenchymal transition-related protein expression in cervical cancer

Epithelial-mesenchymal transition (EMT) plays an important role in cancer invasion and metastasis by enabling cancer cells to depart from the primary tumor, invade surrounding tissue and disseminate to distant organs. The existence and function of EMT in cervical cancer is poorly understood. Placental growth factor (PLGF) has been shown to associate with EMT in various cancers. However, whether PLGF is involved in EMT in cervical cancer remains unclear. Thus the present study examined the relationship between PLGF expression and EMT-related proteins in 110 cervical lesions samples. We detected that PLGF was expressed in 61.8% cervical lesion sections. In addition, PLGF expression is positively correlated with low expression level of E-cadherin and high expression level of vimentin. Serum samples and cervical lavage samples were collected from patients with pre-invasive and invasive lesion of uterine cervix or normal control group, the PLGF levels were determined by enzyme-linked immunosorbent assay (ELISA). We found that a significantly high level of PLGF could be detected both in serum and vaginal lavage compared with normal women group, and there is no significant difference between serum and lavage in PLGF level. In addition, whatever in lavage or in serum, the PLGF level in stage I and II was significantly higher than it in CINIII or cancer in situ. However, there is no significant difference between the stage I and stage II; we also found that exogenous PLGF promotes molecular changes of epithelial-mesenchymal transition (EMT) in siha cells. In addition, application of a specific EKR1/2 inhibitor could reverse the effects of PLGF. These findings suggested that PLGF could regulate the expression of EMT-related proteins and promote migration of siha cells through ERK/MAPK signaling pathway. Therapies that targets PLGF/Flt-1/ERK/MAPK signaling pathway may be beneficial in treatment of cervical cancer.     

血管内皮细胞生长因子在肥胖乳腺癌患者乳腺癌组织中的表达及临床意义

 摘要：　目的 探讨肥胖乳腺癌患者乳腺癌组织中血管内皮细胞生长因子（vascular endothelial growth factor,VEGF）的表达及临床意义。方法 采集45例肥胖乳腺癌患者,37例正常体重乳腺癌患者,用RT-PCR和免疫组织化学检测肥胖组和正常体重组乳腺癌组织VEGF的mRNA和蛋白表达。结果 肥胖组乳腺癌组织VEGF的mRNA和蛋白表达均高于正常体重组 (P<0.05),并且VEGF mRNA和蛋白表达之间呈正相关(r=0.785,P<0.05);VEGF在肥胖乳腺癌组织中的表达与组织学分级、5年复发转移有关,与患者年龄无关。结论 VEGF在肥胖乳腺癌患者中具有高表达趋势,提示VEGF可能在肥胖相关性乳腺癌的发生、演变及预后中起重要作用。     

Epithelial–mesenchymal transition markers in malignant ovarian germ cell tumors

The purpose of this study was to determine the expression and potential clinical role of epithelial‐to‐mesenchymal transition (EMT)‐related factors in malignant ovarian germ cell tumors (MOGCT). Protein expression of E‐cadherin, N‐cadherin, P‐cadherin, Zeb1, HMGA2, and vimentin by immunohistochemistry was analyzed in 42 MOGCT from patients treated in Norway during the period 1981–2001. Expression was analyzed for association with clinicopathologic parameters. E‐cadherin (p = 0.016) and HMGA2 (p = 0.002) expression was significantly higher in immature teratomas and yolk sac tumors compared with dysgerminomas. Vimentin (p < 0.001) and Zeb1 (p = 0.029) staining was significantly higher in immature teratomas compared with yolk sac tumors and dysgerminomas, whereas no significant differences were observed for N‐cadherin and P‐cadherin. EMT‐associated markers were not significantly related to clinicopathologic parameters including age, tumor diameter, and FIGO stage. In conclusion, based on this limited series, EMT‐associated markers are not associated with clinical parameters in MOGCT, in contrast to ovarian carcinoma. EMT‐related proteins are differentially expressed among various MOGCT subtypes, suggesting differences in biological characteristics associated with invasion and metastasis.