ObjectiveTo explore the effect of small interference RNA (siRNA) on the metastasis and invasion of A549 cells after silence the E-cadherin gene (CDH1).MethodsThe CDH1 gene in A549 cells was silenced by siRNA technology, and RT-QPCR and Western blot were used to detect the gene expression and protein expression after silencing. The cells were divided into control group (Control), siRNA negative group (siRNA-NC) and siRNA gene silencing group (siRNA-CDH1). CCK-8 detected cell viability after the gene silencing in 12 h, 24 h, 48 h and 72 h, and cell scratch test to detect cell migration ability. Transwell chamber was used to detect cell invasiveness, and Western blot method was used to detect the expression level of key factors which involved in cell migration, invasion, such as vimentin, matrix metalloproteinases (MMP-2 and MMP-9), and N-cadherin. RT-QPCR was employed to detect the mRNA expression of vimentin, MMP-2, MMP-9, Slug and Snail.ResultsAfter CDH1 silencing, the activity of A549 cells increased significantly and was time-dependent, compared with Control group and siRNA-NC group (P<0.05). The cell scratch test and Transwell chamber test showed that the cell migration and invasion ability increased significantly after CDH1 silencing, compared with Control group and siRNA-NC group (P<0.05). The expression of vimentin, MMP-9, MMP-2 and N-cadherin in siRNA-CDH1 group increased significantly. The mRNA expression level of vimentin, MMP-9, MMP-2 and Slug and Snail also increased. Compared with Control group and siRNA-NC group, the difference was statistically significant (P<0.05).ConclusionCDH1 plays an important role in tumor proliferation and invasion, and its deletion can increase the proliferation and invasion ability of A549 cells.
ObjectiveTo explore the expression of MSI2 in gastric cancer and its association with clinical significance. MethodsThe expression level of MSI2 mRNA in gastric cancer tissues and para-carcinoma tissues was detected by Real-time PCR to explore its clinical significance. The expression level of MSI2 protein was detected by Western blotting. The prognosis of patients with the ratio of MSI2 mRNA expression level in gastric cancer tissues and adjacent normal tissues was more than 2 times and below 2 times were compared. ResultsThere was no significant difference between the expressions of MSI2 mRNA in cancer tissues and para-carcinoma tissues(P > 0.05). The expression level of MSI2 mRNA were associated with the invasion depth (P=0.017), TNM stage (P=0.028), differentiation (P=0.020), and size (P=0.030) of tumor, but no association with other clinical factors such as gender, age, and location were found. The overall survival of the patients with high expression of MSI2 mRNA was significantly shorter than that of the patients with low expression of MSI2 mRNA (χ2=4.221, P=0.040). ConclusionMSI2 expression is associated with the gastric cancer invasion, TNM stage and differentiation, and the patients with higher expression of MSI2 mRNA have poor prognosis, which makes it possible to be a potential therapeutic target.
ObjectiveTo explore the influence mechanism of proliferation and invasion in colon cancer cell after silence of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene. MethodsRT-PCR or Western blot method was used to detect the expression of PTEN mRNA or protein among four colon cancer cell lines (HT-29, WiDr, CaCo-2, and Colo320 cell lines). small interfering RNA (siRNA) was used to synthetize PTEN siRNA and transfect it into colon cancer cells. The expression of PTEN protein after transfecting was detected by Western blot. WsT-1 and invasion assay were used to examine the effects of PTEN siRNA silence on proliferation and invasion in colon cancer cells. ResultsPTEN mRNA and protein were expressed in all the four colon cancer cell lines. After PTEN siRNA transfected into the colon cancer cells, the expressions of PTEN proteins were inhibited in all the four colon cancer cell lines (P < 0.01), and the proliferation and invasion of colon cancer cells were enhanced significantly (P < 0.01). ConclusionsPTEN siRNA play an important role in metastasis process of colon cancer via enhanced its proliferation and invasion. Therefore, the understanding biologic mechanisms for regulation of PTEN might enable better molecular target therapy of treating the colon cancer patients with metastasis.
【Abstract】ObjectiveTo study the effect of down-regulation of E-cadherin on the invasion ability of tumor cells. MethodsHuman pancreatic carcinoma cell line JHP-1 was treated with E-cadherin antisense oligodeoxynucleotied (ASODN). The immunocytochemistry, Western blot were used to detect the expression and the contents of E-cadherin in the tumor cells, and the invasive ability of tumor cells were evaluated by invasive-MTT assay. Results Treated with E-cadherin ASODN, the expression of E-cadherin on JHP-1 cells were reduced, and the protein contents were decreased as well compared with control groups and ODN group. The invasive ability of JHP-1 cells to the basement membrane was increased (P＜0.001) compared with ODN group and control group. ConclusionE-cadherin was related to the invasive ability of tumor cells.
Objective To study the expressions of Wnt5a, MMP2, and MMP14 in the primary lesions of gastric cancer and the influences on clinicopathologic features. Methods The expressions of Wnt5a, MMP2, and MMP14 in the specimens of 106 patients with gastric cancer and 39 patients from the adjacent normal gastric tissues were detected by immunohistochemical staining, χ2 test and non-parametric test were used to analyze the relationships among them and between them and their influences on the clinicopathologic features. Results Extensive expressions of Wnt5a, MMP2, and MMP14 were demonstrated in the gastric cancer, which were significantly higher than those in the normal gastric tissues respectively (Plt;0.05). Positive expression of Wnt5a was associated with larger tumor diameter, deeper depth of invasion, higher degree of regional lymph node metastasis, later TNM stage, and higher rate of lymph node metastasis (Plt;0.05). In addition, Wnt5a expression was also associated with lymphatic infiltration and vascular infiltration (Plt;0.05). The expressions of MMP2 and MMP14 were associated with lymphatic infiltration, but not with vascular infiltration. Higher expressions of MMP2 and MMP14 were correlated with deeper tumor invasion, higher degree of regional lymph node metastasis, later TNM stage, and higher rate of lymph node metastasis (Plt;0.05). In addition, higher expression of MMP2 possesed greater tumor diameter (Plt;0.05). Spearman rank correlation analysis revealed the positive relation between Wnt5a and MMP2 （rs=0.240, P=0.014）, Wnt5a and MMP14 （rs=0.251, P=0.010）, as well as MMP2 and MMP14 （rs=0.444, P=0.000）. Conclusion Higher expressions of Wnt5a, MMP2, and MMP14 seem to promote invasion and metastasis of gastric cancer, and there are positive relations among their expressions.
ObjectiveTo study the expression of matrilysin in gastric cancer and to evaluate the correlation between its expression and invasion, metastasis, and prognosis. MethodsA total of 52 patients with gastric cancer were selected and followed up. The expressions of matrilysin in gastric primary focus, normal gastric mucosa, and metastatic lymph nodes were examined by reverse transcriptionpolymerase chain reaction (RT-PCR), Western blot, and immunohistochemistry, respectively. The correlations between matrilysin expression and tumor invasion, metastasis, and prognosis were assessed. ResultsThe expressions of matrilysin in gastric primary focus and metastatic lymph nodes significantly increased, while decreased or loss in normal gastric mucosa (Plt;0.001). The higher concordance was seen between the levels of mRNA and protein (Plt;0.001). Among patients with infiltrating type, penetrated serosa, area of serosa involved more than 20 cm2, and metastatic lymph nodes more than 7, the expression of matrilysin was significantly higher (Plt;0.01). The survival rate of patients with matrilysin higher expression (34.1%) was significantly lower than that with matrilysin lower expression (55.6%), χ2=9.778， P=0.002. Conclusions Up-regulated expression of matrilysin plays an important role in tumor invasion, metastasis, and poor prognosis, and it is a good molecular marker to reflect the biological behaviors of gastric cancer.
【Abstract】Objective To study the effects of exogenous hyaluronidase on invasive and angiogenic potential of human breast cancer cell line ZR-75-30.MethodsThere were two groups in the study: the study group (hyaluronidase group) and the control group. The invasive potential and the angiogenic potential of human breast cancer cell ZR-75-30 were detected by the invasive model in vitro and technique of double-chamber co-culture that human breast cancer cell line ZR-75-30 and human umbilicus vein endothelium cell ECV-304 were co-cultured. ResultsThe penetrating number of tumor cell in the study group (70.625±11.64) was significantly higher than that in the control group (22.125±6.09),P＜0.01. The tube number from ECV-304 cell induced by ZR-75-30 cell in the study group (34.5±2.4) was significantly higher than that in the control group (8.5±1.5)， P＜0.01. ConclusionExogenous hyaluronidase can reinforth the invasive and angiogenic ability of breast cancer cells.
【 Abstract 】 Objective Overexpressions of epidermal growth factor (EGF) and EGF receptor have been associated with progression and invasive phenotype of pancreatic cancer. However, the underlying molecular mechanism by which EGF worked in pancreatic cancer cells has not been completely understood. In this study, effect of EGF on the invasion and metastasis of pancreatic cancer cells and its regulatory mechanism were investigated. Methods The effects of EGF on the proliferation, adhesion and invasion of pancreatic cancer cells were detected by WST-1 proliferation assay, adhesion assay and invasive assay, respectively. The activity and expression of MMP-2 and MMP-9 were examined by zymography, Western blot and RT-PCR, respectively. The activity of NF- κ B was examined by EMSA. Results EGF could significantly promote the invasiveness of pancreatic cancer cells but did not affect cell proliferation or adhesion. The expressions of NF- κ B and MMP-9 were significantly increased by EGF, but EGF did not affect the activity and expression of MMP-2. Furthermore, EGF stimulated the NF- κ B binding activity. Pretreatment with NF- κ B inhibitors, pyrrolidine dithiocarbamate (PDTC), could significantly inhibit the activity of NF- κ B induced by EGF. Meanwhile, the EGF-induced expression and activity of MMP-9, as well as cell invasiveness were also inhibited by NF- κ B inhibitor. Conclusion EGF could increase the expression and promote the invasiveness of MMP-9 via the activation of NF- κ B in pancreatic cancer cells, which implies that NF- κ B inhibitant, such as PDTC, may diminish the invasiveness of pancreatic cancer cells.
ObjectiveTo investigate the influence of hypoxia on pro-metastasis induced by epithelial-mesenchymal transition (EMT) of human lung adenocarcinoma. MethodsThe human lung cancer cell line H460 was cultured in hypoxic condition and the morphologic changes of the cells were observed under the microscope. The EMT-related markers including E-cadherin and vimentin were detected by Western blot. Transwell migration assay and transwell invasion assay were employed to detect the migratory and invasive activity of cancer cells. ResultsHypoxic induced morphological changes were consistent with the mesenchymal phenotype, such as an elongated fibroblastic morphology, and conversion from a tightly packed epithelial cobblestone pattern to a loosely packed scattered phenotype. Compared with the control group, hypoxic attenuated the quantity of E-cadhenrin, but increased vimentin, which resulted in promotion of migration and invasion of H460. ConclusionHypoxia induces EMT in H460 and enhances the invasive and migratory abilities of lung cancer cells.
Objective To detect the tissue factor (TF) mRNA expression in hepatocellular carcinoma and to elucidate its significance. Methods TF mRNA was detected by reverse transcription-polymerase chain reaction （RT-PCR） in 27 cases of human hepatocellular carcinoma tissue specimen with their adjacent tissues and in 27 non-tumorous process tissues. Then the relationship between mRNA expression and pathological data were analyzed. Results The expression and the relative expression intensity of TF in hepatocellular carcinoma tissues were 62.96%（17/27） and 0.567±0.268 respectively, which were significantly higher than those in their adjacent tissues 〔33.33%(9/27), 0.469±0.184〕 and in 27 non-tumorous process tissue 〔29.63%(8/27), 0.353±0.121〕, Plt;0.05. The relative expression intensity of TF were associated with tumor size, intrahepatic and extrahepatic metastasis and portal vein invasion, but unrelated to gender, AFP level, differentiation, HBsAg, cirrhosis, number of tumor lesions, and lymph node metastasis (Pgt;0.05). Conclusion Expression of TF mRNA were significantly higher in hepatocellular carcinoma and in the invasive and metastatic tissue, which indicated that TF may play an important role in carcinogenesis, invasion and metastasis of hepatocellular carcinoma.