Expression and biological function of N-myc down-regulated gene 1 in human cervical cancer

The expression of N-myc down-regulated gene 1 (NDRG1) has previously been reported to be involved in the proliferation,differentiation,invasion and metastasis of cancer cells,but its role in cervical cancer is still unclear.This study aimed to investigate the expression of NDRG1gene in human cervical cancer and its effect on aggressive tumor behaviors.The NDRG1 expression in cervical tissues and cells was detected by RT-PCR.Specific expression plasmid pEGFP-N1-NDRG1-GFP was used to enhance the expression of NDRG1 in human cervical cancer cell lines.The mRNA and protein level of NDRG1 was assessed by RT-PCR and Western blotting,respectively.Its effects on cell proliferation,migration,invasion,cell cycle and apoptosis were detected by MTT,transwell migration assay and flow cytometry (FCM),respectively.The results showed that the expression of NDRG1 in cervical cancer tissues and cells was significantly lower than in normal cervical tissues (P<0.001).After transfection with pEGFP-N1-NDRG1-GFP,the mRNA and protein expression of NDRG1 was up-regulated in Siha cells,which suppressed cell proliferation (P<0.001),induced cell cycle arrest (P<0.05),reduced invasion and migration of Siha cells (P<0.05),but caused no cell apoptosis.Moreover,vascular endothelial growth factor (VEGF),a tumor-induced angiogenesis factor,was markedly reduced and E-cadherin,a cell adhesion molecule,was increased in the cells transfected with pEGFP-N1-NDRG1-GFP.It was concluded that up-regulated NDRG1 may play a role in the suppression of malignant cell growth,invasion and metastasis of human cervical cancer.     

Silencing MTA1 by RNAi Reverses Adhesion,Migration and Invasiveness of Cervical Cancer Cells （SiHa） via Altered Expression of p53, and E-cadherin/β-catenin Complex

It has been reported that metastasis-associated gene 1 (Mta1) is overexpressed in many malignant tumors with high metastatic potential. In addition, some studies indicated that MTA1 participated in invasion, metastasis, and survival of cancer cells by regulating cell migration, adhesion and proliferation. But the role of MTA1 is unclear in vitro in the development of cervical cancer cells. This study investigated whether and how MTA1 mediated cell proliferation, migration, invasion and adhesion in cervical cancer. MTA1 expression level was detected by Western blot in two cervical cancer cell lines of different invasion potentials. The effects of MTA1 expression on SiHa cell apoptosis, cycle, proliferation, migration, invasion and adhesion were tested by flow cytometry, MTT, wound-healing assay, Transwell assay and adhesion assay, respectively. The expression levels of p53, E-cadherin, and β-catenin activity were evaluated in untreated and treated cells. The results showed that MTA1 protein expression was significantly higher in SiHa than in HeLa, which was correlated well with the potential of migration and invasion in both cell lines. Furthermore, the cell invasion, migration and adhesion capabilities were decreased after inhibition of MTA1 expression mediated by Mta1-siRNA transfection in SiHa. However, no significant differences were found in cell apoptosis, cycle, and proliferation. In addition, E-cadherin and p53 protein levels were significantly up-regulated, while β-catenin was significantly down-regulated in SiHa transfected with the siRNA. These results demonstrated that MTA1 played an important role in the migration and invasion of cervical cancer cells. It was speculated that the decreased migration and invasion capability by inhibiting the MTA1 expression in the SiHa cell line may be mediated through the altered expression of p53, and E-cadherin/β-catenin complex. MTA1 could serve as a potential therapeutic target in cervical cancer.     

Functional expression of voltage-gated sodium channels Nav1.5 in human breast caner cell line MDA-MB-231

Voltage-gated sodium channels （VGSCs） are known to be involved in the initiation and progression of many malignancies, and the different subtypes of VGSCs play important roles in the metastasis cascade of many tumors. This study investigated the functional expression of Nav1.5 and its effect on invasion behavior of human breast cancer cell line MDA-MB-231. The mRNA and protein expression of Nav1.5 was detected by real time PCR, Western Blot and immunofluorescence. The effects of Nav1.5 on cell proliferation, migration and invasion were respectively assessed by MTT and Transwell. The effects of Nav1.5 on the secretion of matrix metalloproteases （MMPs） by MDA-MB-231 were analyzed by RT-PCR. The over-expressed Nav 1.5 was present on the membrane of MDA-MB-231 cells. The invasion ability in vitro and the MMP-9 mRNA expression were respectively decreased to （47.82±0.53）% and （43.97±0.64）% （P〈0.05） respectively in MDA-MB-23 t cells treated with VGSCs specific inhibitor tetrodotoxin （TTX） by blocking Navl.5 activity. It was concluded that Navl.5 functional expression potentiated the invasive behavior of human breast cancer cell line MDA-MB-231 by increasing the secretion of MMP-9.     

Expression of Pinl and Ki67 in Cervical Cancer and Their Significance

In order to investigate the expression levels of Pinl mRNA and protein in cervical cancer and its association with Ki67 and their clinical significance, amplification of Pinl gene was examined by RT-PCR, and the expression of both Pinl and Ki67 protein was detected by immunohistochemistry in cervical cancer tissues. It was shown that the expression levels of Pinl were higher in cervical cancer than in normal cervical tissues （P〈0.05）. The expression of Pinl protein was increased progressively along with the disease process from normal cervix to CIN and to cervical cancer （P〈0.05）. No significant difference in the Pinl expression was found between disease stages （FIGO）, pathological grades or pelvic lymph node metastasis status （P〉0. 05）. The expression of Pin1 was significantly higher in adenocarcinoma than in squamous carcinoma of the uterine cervix （P〈0.05）. In cervical cancer, the overexpression of Pinl was positively correlated with that of Ki67 （P〈 0. 05）. These results suggested that the overexpression of Pinl was closely related with cancer cell proliferation or progression of cervical cancer and contributed to oncogenesis. Pinl may serve as a potential marker for cervical cancer diagnosis.     

Vascular Endothelial Growth Factor165-regulated Nasopharyngeal Carcinoma Cell Lines Invasion and Migration Involve Expression and Activation of Matrix Metalloproteinase-2

The effect of vascular endothelial growth factor （VEGF） overexpression on matrix metalloproteinase-2 （MMP-2） in nasopharyngeal carcinoma （NPC） cells in vitro and the possible mechanism involved were investigated, and the correlation between the expression of VEGF and MMP-2 in NPC evaluated. The NPC cells were transfected with PAd-trackVEGF165 plasmid. The expression levels of VEGF and MMP-2 mRNA and protein in NPC cells were detected by semi-quantitative RT-PCR and Western blot respectively. It was found that the expression of VEGF and MMP-2 mRNA and protein was significantly increased in NPC cells after transfection of VEGF 165. It was concluded that the expression of VEGF was correlated to the in vitro invasion of NPC cells, and the induction of MMP-2 by VEGF was a key process of NPC cell invasion.     

Hypoxia-inducible factor-1α and semaphorin4D genes involved with tumor-associated macrophage-induced metastatic behavior and clinical significance in colon cancer

Background Hypoxia promotes tumor angiogenesis and hypoxia-inducible factor-1 alpha （HIF-lg） plays a pivotal role in this process. Recently identified pro-angiogenic factor, semaphorin4D （Sema4D） also promotes angiogenesis and enhances invasive proliferation in some tumors. Furthermore, tumor-associated macrophages （TAMs） can increase the expression of HIF-la and Sema4D in cancer cells and thus influence tumor growth and progression. The purpose of this study was to evaluate the effect of TAMs on the expression of Sema4D and HIF-la and the impact of biologic behavior in colon cancer cells. Methods Immunohistochemistry was used to analyze HIF-la and Sema4D expression in 86 curatively resected colon cancer samples and 52 normal colon tissues samples. The relationship between their expression and clinicopathological factors was analyzed. Furthermore, macrophage-tumor cell interactions, such as metastasis, angiogenesis, were also studied using in vitro co-culture systems. Statistical analysis was performed using SPSS 17.0 software （SPSS Inc., USA）. Differences between two groups were analyzed with Student＇s t test. Results HIF-la （58%） and Sema4D （60%） were expressed at a significantly higher level in tumors than in normal tissues （P 〈0.01, for both）. Furthermore, HIF-la and Sema4D expression was significantly correlated with lymphatic metastasis, specific histological types and TNM stages （P 〈0.05）, but not with age and tumor size （P 〉0.05）. Sema4D expression was correlated with that of HIF-la （r=0.567, P 〈0.01）. TAMs markedly induced HIF-la and Sema4D expression in colon cancer calls and subsequently increased their migration and invasion. Conclusions HIF-la and Sema4D expression are closely related to lymphatic metastasis, specific histological types and TNM stages in colon cancer. Furthermore, TAMs promote migration and invasion of colon cancer cells and endothelial tube formation, possibly through up-regulation of HIF-la and Sema4D.     

Inhibitory effects of microRNA-34a on cell migration and invasion of invasive urothelial bladder carcinoma by targeting notch1

MicroRNAs (miRNAs or miRs) are a class of short, non-coding RNAs that participate in various oncological processes. This study aims to explore the roles of microRNA-34a (miR-34a) in invasive urothelial bladder carcinoma. miR-34a was transfected into bladder cancer cell lines 253J and J82. The miR-34a expression levels in tissues and cells were detected by using qRT-PCR. The Notch1 expression was detected by qRT-PCR and Western blotting. Cell migratory and invasive abilities were measured by Transwell chamber assay. Bioinformatics and luciferase assay were performed to predict and analyze the binding sites between miRNA-34a and Notch1. It was found that there was aberrant expression of miR-34a in bladder cancer tissues. Moreover, we revealed that ectopic expression of miR-34a suppressed cell migration and invasion, while forced expression of Notch1 increased cell migratory and invasive abilities. Finally, we observed that miR-34a transfection significantly down-regulated luciferase activity and reduced the mRNA and protein levels of Notch1. Our study concluded that microRNA-34a antagonizes Notch1 and inhibits cell migration and invasion of bladder cancer cells, which indicates the tumor-suppressive function of microRNA-34a in bladder cancer.     

Human ovarian neoplasm cell CD147 stimulates production and activation of matrix metalloproteinases in co-cultures with mouse fibroblasts

Objective: To investigate the expression of CD147 on human ovarian neoplasm cell lines and its influence on production and activation of matrix metallproteinases (MMPs). Methods : The expression of CD147 on different human ovarian neoplasm cell lines was studied by western blotting. Co culture was carried out to investigate the stimulative effect of the positive expression CD147 cell HO-8910 on the production of MMPs of fibroblast cell in vitro. Zymography and immune blotting were used to study the production and activity of positive MMPs, at the time, to explore the relation between CD147 and MMPs. Results: CD147 was positively presented in 2 ovarian neoplasm cell lines (HO-8910, 3-AO), but in SKOV3, TC-1,NIN3T3 cell was negative. MMP-2 and MMP-9 were detected by HO-8910 cell line, mouse fibroblast cell and co-culture cells ； but the expression in co-culture cell is obviously higher than individual cultures of each type alone. CD147 stimulated MMPs in dose-dependent manner. Conclusion: CD147 causes increased production and activation of MMP-2, MMP-9. CD147 is probably a indirect marker of some ovarian cancer cells with invasion and metastasis.     

Expression and Clinical Implication of HIF-1α and VEGF-C in Non-small Cell Lung Cancer

To study the expression and implication of HIF-1α and VEGF-C in non-small cell lung cancer （NSCLC） and its relationship with clinical pathological features of NSCLC, immunohisto-chemical SP was used to detect the expression of HIF-1α and VEGF-C proteins in 48 NSCLC tissues and the same para-cancerous tissues. The positive rates of HIF-1α and VEGF-C were 70.8% （34/48） and 68.8% （33/48） respectively. The expression of HIF-1α protein was detected in a significantly greater proportion in NSCLC carcinoma tissues than that in para-cancerous tissues （12.5% and 16.7%, P〈0.05）. The positive rates of HIF-1α and VEGF-C were correlated with lymph node metastasis and TNM stage. No relationship was found between the two factors and age, sex, pathological subtypes and histological grades. The positive rates between HIF-1α and VEGF-C were correlated （P〈0.05）. HIF-1α and VEGF-C were over-expressed in NSCLC. They may be involved in the carcinogenesis of NSCLC, and play an important role in invasion and metastasis of NSCLC. HIF-1α and VEGF-C work synergically in the process of NSCLC.     

Vascular Endothelial Growth Factorl65-regulated Nasopharyngeal Carcinoma Cell Lines Invasion and Migration Involve Expression and Activation of Matrix Metalloproteinase-2

The effect of vascular endothelial growth factor (VEGF) overexpression on matrix met-alloproteinase-2 (MMP-2) in nasopharyngeal carcinoma (NPC) cells in vitro and the possible mechanism involved were investigated, and the correlation between the expression of VEGF and MMP-2 in NPC evaluated. The NPC cells were transfected with PAd-trackVEGF165 plasmid. The expression levels of VEGF and MMP-2 mRNA and protein in NPC cells were detected by semi-quantitative RT-PCR and Western blot respectively. It was found that the expression of VEGF and MMP-2 mRNA and protein was significantly increased in NPC cells after transfection of VEGF165. It was concluded that the expression of VEGF was correlated to the in vitro invasion of NPC cells, and the induction of MMP-2 by VEGF was a key process of NPC cell invasion.     

Investigation of tumor necrosis factor receptor-p55 expression in human colorectal cancer

Objective: To investigate the clinical significance of tumor necrosis factor receptor-p55 (TNFR-p55) expression and its relationship with the clinical pathology and Dukes‘ classification of human colorectal cancer. Methods: SABC immunohistochemistry method was used to examine TNFR-p55 expression in 91 specimens of colorectal cancer, 81 surrounding mucosas of the tumors and 13 normal tissues. Results: TNFR-p55 expression in colorectal cancer was significantly higher than that in the surrounding mucosa andnormal tissues, and it was significantly higher in the surrounding mucosa than in normal tissue. TNFR-p55 was inversely correlated to serous membrane invasion and lymph node metastasis of colorectal cancers.TNFR-p55 expression in Dukes‘ A and B stages was significantly higher than that of C stage. Conclusion: TNFR-p55 expression was important to determine the degree of malignancy and assess invasion depth, lymph node metastasis and Dukes‘ classification in colorectal cancer.     

The Effect of RhoC siRNA on the Invasiveness and Proliferation of Human Cervical Cancer Cell Line SiHa Cells

This study investigated the effect of RhoC GTPase on the proliferation and metastasis of cervical cancer cells, SiHa cells, in vitro. RhoC siRNA was introduced into SiHa cells to silence the RhoC gene. The mRNA and protein expression of RhoC, before and after RhoC siRNA transfection, was examined by RT-PCR and Western blotting, respectively. The proliferation and apoptosis of SiHa cells were examined by MTT assay and flow cytometry （FACS）, respectively. Adhesive rate was evaluated by Matrigel adhesive assay, and the invasive capability and migration capability were assessed by transwell invasive assay and migration assay, respectively. The results showed that after the RhoC siRNA transfection, the mRNA and protein expression of RhoC was down-regulated in SiHa cells. The down-regulation of RhoC GTPase did not affect the cell proliferation and apoptosis （P〉0.05）, but it did suppress SiHa cells＇ adhesion to matrigel （P〈0.01）, the invasive capability （P〈0.01） and the migration capability （P〈0.01）. It was concluded that RhoC obviously promotes the adhesion, invasion and migration of SiHa cells in vitro, but not proliferation and apoptosis, suggesting that RhoC plays an important role in the progression in cervical cancer.     

Comparison of hyaluronidase expression,invasiveness and tubule formation promotion in ER （-） and ER （＋） breast cancer cell lines in vitro

Background Hyaluronidase （Hyase） is an enzyme which hydrolyses hyaluronan （HA）, a large nonsulfated glycosaminoglycan. Several genes have been identified to code for hyaluronidases in humans. Its role has only recently been underlined in the invasion of prostate cancer, colonic cancer, and breast cancer. Moreover, the findings were in agreement with some experimental results which showed that HA-derived oligosaccharides had angiogenesis-promoting activity. All these findings prompted us to investigate factors that had been characterized as putative invasive factors in different human breast cancer-derived cell lines. Methods We selected two series of human breast cancer-derived cell lines whose expression of estrogen receptors （ER） was previously published. Hyaluronidase secretion in culture medium and expression of matrix metallo-proteinase （MMP）-9, cathepsin-D （cath-D） and vascular endothelial growth factor （VEGF） by cells were determined. We also investigated cell invasiveness in the Matrigel invasion assay, and studied the capability of cancer cells to promote in vitro formation of tubules by endothelial cells. Results ER（-） cells secreted significantly more hyaluronidase （P 〈0.001） and expressed significantly more VEGF （P 〈0.01）, MMP-9 （P 〈0.05） and cath-D （P 〈0.0001） than ER（＋） cells. Invasion through Matrigel by ER（-） Hyase（＋） cells was significantly higher than that by ER（＋） Hyase（-） cells （P 〈0.05）. In both cases, invasion was decreased by heparin （P 〈0.05）. When ECV-304 endothelial cells were co-cultivated in millicell chambers with cancer cells, ECV-304 cells were induced to form tubules. Tubule formation was demonstrated to be more prominent with ER（-） Hyase（＋） cells than with ER（＋） Hyase（-） cells （P 〈0.05）. Conclusion Invasive features of ER（-） breast cancer cells can be characterized in vitro by an invasive Matrigel assay, as the induction of tubule formation by ECV-304 endothelial cells, higher secretion of hyaluronidase, and higher expression of proteinases MMP-9, cath-D, and the angiogenesis promoting factor VEGF.     

Implication of EMT Induced by TGF-β1 in Pancreatic Cancer

This study examined the implication of EMT induced by TGF-β1 in pancreatic cancer invasion. TGF-β1 expression was determined in 29 cases of human pancreatic carcinoma （PC） by immunohistochemistry and the results were compared with those of pathological examination. Moreover, the effects of TGF-β1 on the phenotype and invasion of pancreatic cancer cell line Panc-1 were also investigated. TGF-β1 was detected in 12 cases （41.4 %） of PC. Significant correlation was found between the expression of TGF-β1 and lymph node involvement （P=0.047） and the depth of invasion （P=0.035）. TGF-β1 obviously promoted EMT of Panc-1 cell lines and their invasion ability was substantially enhanced. TGF-β1 may promote the malignancy of pancreatic cancer by triggering EMT.     

非小细胞肺癌组织基质金属蛋白酶-3和组织金属蛋白酶抑制剂-3的表达

目的 探讨非小细胞肺癌(NSCLC)组织中基质金属蛋白酶-3(MMP-3)和组织金属蛋白酶抑制剂-3(TIMP-3)的表达及其意义.方法 采用免疫组化S-P法检测56例非小细胞肺癌组织和35例癌旁正常肺组织中MMP-3和TIMP-3的表达.结果 非小细胞肺癌组织中MMP-3的表达显著高于癌旁正常肺组织(P<0.05).非小细胞肺癌中MMP-3的表达,有淋巴结转移者高于无淋巴结转移者(P<0.05),Ⅲ+Ⅳ期高于I+Ⅱ期(P<0.05).非小细胞肺癌组织中TIMP-3的表达显著高于癌旁正常肺组织(P<0.05),非小细胞肺癌中TIMP-3的表达有淋巴结转移者低于无淋巴结转移者(P<0.05).结论 MMP-3和TIMP-3表达与非小细胞肺癌的侵袭、转移密切相关. Abstract： Objective To investigate the expression of MMP-3 and TIMP-3 in non-small cell lung cancer tissue and their significance. Methods The expression of MMP-3 and TIMP-3 in 56 lung carcinoma tissues and 56 para-cancerous normal lung tissues were detected by immuno histochemishy tecneque. Results The expression of MMP-3 in non-small cell lung cancer tissues was significantly higher than that in para-cancerous normal lung tissues (P<0.05).The expression of MMP-3 was significantly higher in non-small cell lung cancer with lymph node metastasis than that in lung carcinomas without lymph node metastasis (P<0.05),and was significantly higher in stage-Ⅲ and stage-IV tumours than that in stage-I and stage-Ⅱtumours (P<0.05).The expression of TIMP-3 in non-small cell lung cancer tissues was significantly higher than that in para-cancerous normal lung tissues (P<0.05).The expression of TIMP-3 was significantly lower in non-small cell lung cancer with lymph node metastasis than that in lung carcinomas without lymph node metastasis(P<0.05). Conclusions The expression of MMP-3 and TIMP-3 is closely correlated with invasion and metastasis of non-small cell lung cancer.     

Inhibitory effect of ginsenoside Rg3 on ovarian cancer metastasis

Background Ginsenosides are main components extracted from ginseng, and ginsenoside Rg3 is one of the most important parts. Ginsenoside Rg3 has been found to inhibit several kinds of tumor growth and metastasis. The present study was undertaken to investigate the effect of ginsenoside Rg3 on human ovarian cancer metastasis and the possible mechanism. Methods The experimental lung metastasis models of ovarian cancer SKOV-3 and the assay of tumor-induced angiogenesis were used to observe the inhibitory effects of Rg3 on tumor metastasis and angiogenesis. The effect of Rg3 on invasive ability of SKOV-3 cells in vitro was detected by Boyden chamber, and immunofluorescence staining was used to recognize the expression of matrix metalloproteinase 9 （MMP-9） in SKOV-3 cells. Results In the experimental lung metastasis models of ovarian cancer, the number of tumor colonies in the lung and vessels oriented toward the tumor mass in each ginsenoside Rg3 group, was lower than that of control group. The invasive ability and MMP-9 expression of SKOV-3 cells decreased significantly after treatment with ginsenoside Rg3. Conclusions Ginsenoside Rg3 can significantly inhibit the metastasis of ovarian cancer. The inhibitory effect is partially due to inhibition of tumor-induced an qioqenesis and decrease of invasive ability and MMP-9 expression of SKOV-3 cells.     

Expression and Clinical Implication of HIF-la and VEGF-C in Non-small Cell Lung Cancer

To study the expression and implication of HIF-la and VEGF-C in non-small cell lung cancer (NSCLC) and its relationship with clinical pathological features of NSCLC, immunohistochemical SP was used to detect the expression of HIF-1а and VEGF-C proteins in 48 NSCLC tissues and the same para-cancerous tissues. The positive rates of HIF-1а and VEGF-C were 70.8% (34/48) and 68.8% (33/48) respectively. The expression of HIF-la protein was detected in a significantly greater proportion in NSCLC carcinoma tissues than that in para-cancerous tissues (12.5% and 16.7%,P<0.05). The positive rates of HIF-1а and VEGF-C were correlated with lymph node metastasis and TNM stage. No relationship was found between the two factors and age, sex, pathological subtypes and histological grades. The positive rates between HIF-1а and VEGF-C were correlated (P<0.05).HIF-1а and VEGF-C were over-expressed in NSCLC. They may be involved in the carcinogenesis of NSCLC, and play an important role in invasion and metastasis of NSCLC. HIF-1а and VEGF-C work synergically in the process of NSCLC.