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Objective To explore the effect of silencing metastasis-associated in colon cancer 1 ( MACC1 ) gene on proliferation and invasion of cervical cancer Hela cells. Methods siRNA MACC1 and siRNA NC were transfected into cervical cancer Hela cells with liposomes. The expression of MACC1 protein and mRNA was detected by western blot and RT-PCR. Cell viability and invasion ability were measured by MTT and transwell assay, respectively. The expression of cyclin D1, cyclin E matrix metalloproteinase 2 (MMP2), MMP9 and the level of extracellular regulated protein kinases ( ERK) phosphorylation was detected by western blot. Results Compared with siRNA NC, the expression of MACC1 protein and mRNA was down-regulated, cell viability and invasion ability were reduced ( P < 0. 01 ) , cell cycle was arrested at G1 phase (P < 0. 01), the expression of cyclin D1, cyclin E, MMP2, MMP9 and p-ERK1/2 was down-regulated (P< 0. 01). Conclusions siRNA MACC1 can inhibit proliferation and invasion of cervical cancer Hela cells, which migiht be related to down-regulation of p-ERK.
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Objective To explore the effect of silencing metastasis-associated in colon cancer 1 ( MACC1 ) gene on proliferation and invasion of cervical cancer Hela cells. Methods siRNA MACC1 and siRNA NC were transfected into cervical cancer Hela cells with liposomes. The expression of MACC1 protein and mRNA was detected by western blot and RT-PCR. Cell viability and invasion ability were measured by MTT and transwell assay, respectively. The expression of cyclin D1, cyclin E matrix metalloproteinase 2 (MMP2), MMP9 and the level of extracellular regulated protein kinases ( ERK) phosphorylation was detected by western blot. Results Compared with siRNA NC, the expression of MACC1 protein and mRNA was down-regulated, cell viability and invasion ability were reduced ( P < 0. 01 ) , cell cycle was arrested at G1 phase (P < 0. 01), the expression of cyclin D1, cyclin E, MMP2, MMP9 and p-ERK1/2 was down-regulated (P< 0. 01). Conclusions siRNA MACC1 can inhibit proliferation and invasion of cervical cancer Hela cells, which migiht be related to down-regulation of p-ERK.
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OBJECTIVE Metastasis-associated in colon cancer-1 (MACC1) is an oncogene that has been newly identified. It promotes tumor proliferation and invasion via the MET pathway. Our study investigated the effects of Saikosaponin-b(SS-b) on the proliferation and apoptosis of HepG2 cells and its regulation on MACC1/c-Met/Akt signaling pathway. METHODS HepG2 cells were treated with SS-b (10-800 g·L-1) for 48 h in vitro. The CCK-8 assay was used to assess cell proliferation, and cell apoptosis was determined by Hoechst33258 staining, AnnexinⅤ/PI staining and caspase 3 assay. RT-PCR was used to examine the expression of MACC1, c- MET and hepatocyte growth factor (HGF) mRNA. MACC1 protein was detected by Western blot and immunohistochemistry. The protein expressions of p-c-MET, c-MET, p-AKT, AKT, p-BAD, BAD were measured by Western blot. RESULTS SS-b inhibited the growth of HepG2 cells in dose-dependent way and induced cell apoptosis significantly. HepG2 cells showed karyopyknosis, fragmentation and fluorescence highlight in SS-b treatment group. FCM results showed that apoptosis rate of HepG2 cells increased with SS- b concentration. The immunofluores?cence results showed that the MACC1 expression decreased significantly in HepG2 cells treated with SS-b. The expression levels of MACC1, c-MET and HGF mRNA in HepG2 cells were significantly inhibited by SS-b. SS-b also significantly decreased the protein expressions of MACC1, p-c-MET and p-AKT while increased the expression of p-BAD and caspase 3 in HepG2 cells(P<0.05). CONCLUSION SS-b inhibited the proliferation and induced the apoptosis of HepG2 cells by targeting the MACC1/c-Met/Akt signaling pathway.
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The protein product of metastasis-associated in colon cancer 1 (MACC1) gene induces the activation of hepatocyte growth factor/mesenchymal-epithelial transition factor (HGF/c-Met) signaling pathway through transcriptionally activating c-Met gene and upregulating its expression to further promote tumor invasion and metastasis.High level expression of MACC1 is associated with the occurrence and metastasis of a wide variety of human tumors, such as colon cancer, gastric cancer, liver cancer, lung cancer, ovarian cancer etc.In addition, the overexpression of MACC1 is also closely associated with clinical TNM stages and distant metastasis.Thus, MACC1 can serve as an independent indicator of tumor metastasis and prognosis, and become a new target for gene therapy.
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Objective To investigate the intratumoral expression of the metastasis-associated in colon cancer 1 (MACC1) and to discuss the clinical relevance associated with hepatocellular carcinoma (HCC). Methods Immunohistochemistry staining was performed to assess the expressions of MACC1 protein in 108 HCC liver tissues and matched adjacent non-tumor liver tissues. Quantilative real-time PCR (qPCR) was employed to examine MACC1 mRNA expression in 30 HCC tissues and the corresponding adjacent non-tumor tissue, and 20 normal liver tissues. The association of MACC1 expression in HCC tissues with the clinicopathological and prognosis was analyzed. Results Immunohistochemistry results showed that the positive rate of MACC1 in HCC tissues was significantly higher than that in the para-carcinoma tissue (49.1% vs 34.2%, P=0.011). qPCR revealed that MACC1 mRNA expression in HCC tissues was significantly higher than that in the noncancerous liver tissues and normal liver tissues(P<0.001). One-factor analysis of variance indicated that MACC1 expression in human HCC tissue was significantly related to tumor size, presence of capsule and pathological types(P<0.05). Survival analysis disclosed that the 1-year, 3-year and 5-year survival rates were 0.650, 0.492, and 0.280 for MACC1 positive patients and 0.799, 0.684, and 0.566 for MACC1 negative patients, respectively, with significant differences found between the two groups(P=0.006). Conclusion Expression of MACC1 in HCC tissues is associated with malignancy evolution and clinical outcome of HCC patients; MACC1 may serve as an important molecule for predicting prognosis patients with hepatic cancer and a new target for gene therapy of liver cancer.
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Objective: To knock down the expression of metastasis-associated in colon cancer-1 (MACC1) gene in drug-resistant ovarian cancer cell line SKOV3/cisplatin (DDP) by RNA interference technology, and to investigate its effect on chemoresistance to DDP. Methods: Ovarian cancer SKOV3/DDP cells were not transfected (blank group, named as SKOV3/DDP cells), or transfected with empty plasmid (negative control group, named as SKOV-3/DDP-shVector cells) or combinant plasmid containing MACC1 specific small hairpin RNA (shRNA) (experimental group, named as SKOV-3/DDP-shMACC1 cells). The mRNA and protein expressions of MACC1 were detected by reverse transcription-PCR (RT-PCR) and Western blotting, respectively. The half inhibition concentration (IC 50) of DDP was determined by MTT assay. The apoptosis was examined by flow cyometry. Results: After transfection of MACC1 shRNA into SKOV-3/DDP cells, the expressions of MACC1 mRNA and protein obviously decreased by (83.39±1.26)% and (82.25±2.94)%, respectively, as compared with those of the blank group (P < 0.05). Compared with the blank group and the negative control group, the value of IC50 of DDP in SKOV-3/DDP-shMACC1 cells significantly decreased [(26.09±0.91) μmol/L vs (47.50±0.40) μmol/L and (47.08±0.45) μmol/L, P < 0.05]. The apoptotic rate of SKOV-3/DDP-shMACC1 cells was significantly higher than those in the SKOV-3/DDP-shVector cells and the SKOV-3/DDP cells [(1 3.77±0.60)% vs (4.23±0.30)% and (3.63±0.30)%, P < 0.05]. Conclusion: The sensitivity to DDP in ovarian cancer SKOV-3/DDP cells can be enhanced by inhibition of MACC1 gene expression induced by RNA interference. Copyright© 2014 by TUMOR.
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Metastasis-associated in colon cancer-1 (MACC1) is a recently discovered gene associated with colon cancer metastasis,there is significant relationship indicated from some studies between MACC1 and different malignant tumors.It may play an important role in the regulation of tumors metastasis.This article reviewed the expression and regulating function of MACC1 in different cancers including colorectal cancer,hepatic cancer,gastric cancer,lung cancer,ovarian cancer,breast cancer,and so on.It may offer clues to find a new target for target treatment of cancer metastasis.
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Objective To examine the expression of metastasis-associated in colon cancer 1 (MACC1) gene in ovarian cancer cell lines and investigate its effect on biological behaviors of ovarian cancer cells.Methods The expression of MACC1 was examined by qRT-PCR and Western blot analysis in four ovarian cancer cell lines inculding OVCAR3,ES-2,SKOV3 and HO-8910.When the MACC1 was transfected to OVCAR3 cells,fluorogenic quantitative PCR was used to filter and identify MACC1 gene after the efficient silencing.Changes of adhesion in the cells were analyzed by an adhesion assay.Transwell migration and invasion assay and in vitro vascular mimicry assay were used to detect migration,invasion and angiogenesis of OVCAR3 cells in vitro.Results The expression of MACC1 gene was higher in OVCAR3 compared to other cell lines.qRT-PCR confirmed that the expression of MACC1 was silenced successfully after transient transfected MACC1-siRNA into OVCAR3 cells.After successful silencing the MACC1 expression,the adhesion ability was inhibited to some degree.In transwell migration assay,the numbers of cells in upper chamber passing through the membrane in transfected group were less than control groups (245.5 ±12.8,500.3±16.5 and 496.3±13.1 respectively),while in transwell invasion assay,the numbers of cells in upper chamber passing through the membrane in transfected group were less than the negative group and control group (185.3±14.1,405.7±9.1 and 416.3±11.5 respectively),both with markedly differences among the three groups.In tube formation assay,the distrubition of HUVECs was diffused with less junctions,and the average number of complete tubular structure was decreased in transfected group compared to the corresponding controls.Conclusion RNA interference inhibits the expression of MACC1 and effectively inhibits the metastasis and invasion abilities of ovarian cancer cells in vitro,and MACC1 is expected to become the target gene of ovarian cancer treatment.
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Objective To explore the expression and clinical relevance of metastasis-associated colon cancer-1 (MACC1) and C-MET proteins in hepatocellular carcinoma (HCC) tissue.Methods The expressions of MACC1 and C-MET were detected in 51 specimens of HCC and paraneoplastic liver tissue,normal liver tissue in 13 healthy cases using immunohistochemistry and Western blotting.The correlations of the expressions of MACC1 and C-MET proteins were evaluated,survival rates were observed,the relationship between the expression of MACC1,C-MET proteins and the clinicopathologic features of HCC were analyzed.Results The positive rate of MACC1 and C-MET proteins was 80.4％ and 76.5％ in HCC tissue,the relative expressions were 0.645 ± 0.047 and 0.504 ± 0.023 respectively,which was significantly different from those in paraneoplastic liver tissue and normal liver tissue (respectively F =173.308,252.817,all P =0.000).The survival analysis showed that the three-year survival rate in patients with positive MACC1 and C-MET expressions was significantly lower than that in patients with negative expressions (respectively x2 =3.934,4.439,all P ＜ 0.05),the positive rate and relative expressions of MACC1 and C-MET were significantly correlated with TNM stage,portal vein cancer thrombus and pathology typing (P ＜ 0.05).Conclusions The expression of MACC1 and C-MET is associated with the malignant progression of HCC.MACC1 may serve as a independent prognostic factor for advanced HCC and a possible therapy target for the treatment of HCC.
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Background and purpose: The metastasis-associated in colon cancer-1 (MACC1) is highly expressed in different cancers and has an effect on the proliferation and apoptosis of tumor cells through the regulation of extracellular signal-regulated kinase (ERK)1/2 pathway. However, the role of MACC1 in ovarian cancer has been rarely studied. The study was aimed to suppress MACC1 gene expression by siRNA and explore the relationship between MACC1 expression and chemosensitivity to cisplatin in ovarian cancer cell line SKOV3/DDP. Methods:Empty plasmid p-super-EGFP-1 (negative control group) and p-super-EGFP-MACC1 shRNA (experimental group) were transfected into ovarian cancer cell SKOV3/DDP respectively. SKOV3/DDP cells without transfection were used as blank group. Then, MACC1 mRNA and protein levels were measured by RT-PCR and Western blot, respectively. Cell proliferation and IC50 of cisplatin was determined by methyl thiazolyl tetrazolium test (MTT). Apoptosis rate was determined by lfow cytometer (FCM). ERK1/2 and p-ERK1/2 protein levels were determined by Western blot. Results:Compared with those in blank and negative control groups, MACC1 mRNA and protein levels deceased in experimental group. The IC50 of cisplatin in experimental group was lower than that in the other groups (26.094 vs 47.501/47.089μmol/L, P<0.05). There was a lower expression of p-ERK1/2 in experimental group (0.3979 vs 00.6712/0.6681, P<0.05). Apoptosis rate was significantly higher in the experimental group before and after treatment of cisplatin (1.32%vs 0.66%/0.48%, P<0.05;36.70%vs 18.53%/16.60%, P=0.000). Conclusion:MACC1 gene may be involved in cisplatin resistance phenomenon in SKOV3/DDP cells through ERK1/2 pathway.
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Metastasis-associated in colon cancer-1 (MACC1),a newly discovered gene which controls tumor growth and metastasis,abnormally expresses in a variety of malignant tumors.As a key regulator of HGF-MET signal pathway,its coding protein can obviously promote invasion and metastasis of tumor cells.