MACC1 mRNA Research Articles

Cantharidin and Its Analogue Norcantharidin Inhibit Metastasis-Inducing Genes S100A4 and MACC1.

Colorectal cancer (CRC) is the third most prevalent and second deadliest cancer worldwide. In addition, metastasis directly causes up to 90% of all CRC deaths, highlighting the metastatic burden of the disease. Biomarkers such as S100A4 and MACC1 aid in identifying patients with a high risk of metastasis formation. High expression of S100A4 or MACC1 and to a greater extent the combination of both biomarkers is a predictor for metastasis and poor patient survival in CRC. MACC1 is a tumor-initiating and metastasis-promoting oncogene, whereas S100A4 has not been shown to initiate tumor formation but can, nevertheless, promote malignant tumor growth and metastasis formation. Cantharidin is a natural drug extracted from various blister beetle species, and its demethylated analogue norcantharidin has been shown in several studies to have an anti-cancer and anti-metastatic effect in different cancer entities such as CRC, breast cancer, and lung cancer. The impact of the natural compound cantharidin and norcantharidin on S100A4 and MACC1 gene expression, cancer cell migration, motility, and colony formation in vitro was tested. Here, for the first time, we have demonstrated that cantharidin and norcantharidin are transcriptional inhibitors of S100A4 and MACC1 mRNA expression, protein expression, and motility in CRC cells. Our results clearly indicate that cantharidin and, to a lesser extent, its analogue norcantharidin are promising compounds for efficient anti-metastatic therapy targeting the metastasis-inducing genes S100A4 and MACC1 for personalized medicine for cancer patients.

Abstract 2879: MACC1 overexpression in colorectal cancer is driven by chromosomal instability and is associated with molecular subtype

Abstract Metastasis-Associated in Colon Cancer 1 (MACC1) is a strong prognostic marker for human colorectal cancers (CRC). The gene is a key regulator of the HGF/c-Met pathway and its overexpression is associated with cancer cell proliferation, migration, invasiveness and metastasis. The contexture of MACC1 overexpression in cancers is still poorly understood. Here we used the data released by The Cancer Genome Atlas (TCGA) and in silico approaches for a deep and integrative analysis of MACC1 expression modalities in CRCs. First, we curated COAD-READ TCGA datasets and obtained full clinical, genomic, and transcriptomic annotation for a total of 259 CRC samples. In tumors, we showed high heterogeneity of MACC1 mRNA expression levels with data varying from 5.58 to 13.07 U (RNAseq log transformed data - RSEM normalized). By correlating MACC1 expression to clinical data, we showed its levels were associated with anatomic regions of the disease (p=6.35.10-5). The tumors from descending colon and rectum having higher levels than those of the ascending colon and caecum (p=7.44-06). High MACC1 was also associated with higher tumor stages (p=0.01) and nodal invasion (p=0.03). At genomic level, MACC1 mutation was not frequent in CRCs (<2%). The gene is however frequently subjected to gene copy number variation (CNV) due to chromosomal instability and polyploidy (MACC1 CNV values > normal in 155/259: 60%). Correlation analysis indicated strong positive association between MACC1 copy number variations and mRNA expression levels (p<10-16). In an independent dataset, we validate that gain of MACC1 gene copy number and associated higher expression levels correlated with metastasis dissemination and worse outcome of the patients. By analyzing MACC1 with respect of CRC subtypes, we confirmed higher MACC1 levels in MSS tumors with high chromosomal instability (CIN) compared to MSI or genomic stable tumors (GS) (p=1.16-14). Regarding association with CMS1-4 transcriptomic subtypes, the highest expression of MACC1 were observed in CMS2-(epithelial), and CMS4 (mesenchymal) as compared with CMS3 (epithelial with metabolic dysregulation) or CMS1 (MMR deficient). The subset of tumors with very low MACC1 expression were mainly those altered for MLH1 (MSI-H- MMR tumors), notably most of them have normal ploidy. Deconvolution analysis of immune contexture revealed negative correlation between MACC1 expression and immune activation. Stratification using MACC1 as prognostic marker within these tumor types will be evaluated and the gene will also be presented with respect of alterations of key cancer genes in these tumors. Overall, the present study gives insight into the contexture of MACC1 expression in CRC. We showed MACC1 overexpression is largely driven by chromosomal instability and DNA copy number gains. Its expression was dependent on the tumor molecular subtypes. Citation Format: Vincent Vuaroqueaux, Hoor Al-Hasani, Dennis Kobelt, Thomas Risch, Susen Burock, Marie-Laure Yaspo, Heinz-Herbert Fiebig, Ulrike Stein. MACC1 overexpression in colorectal cancer is driven by chromosomal instability and is associated with molecular subtype [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2879.

Abstract B14: Activation of the MACC1/PIM/cMyc axis confers resistance to PI3K/mTOR inhibition in PIK3CA mutant NSCLC

Abstract Introduction: The PI3K pathway is often dysregulated in lung cancer, making it an ideal therapeutic target. Pan- and isoform-specific inhibitors of the PI3K pathway are currently being evaluated in clinical trials. However, all patients treated thus far develop progressive disease due to the emergence of bypass signaling mechanisms. We developed a panel of lung cancer preclinical models to elucidate potential mechanisms of resistance to PI3K-mTOR inhibitor GDC-0980 (apitolisib). In-depth characterization of the PIK3CA mutant positive H1975 sensitive cells (H1975P) versus apitolisib-resistant cells (H1975GR) identified activation of MACC1 as an early marker of drug resistance and the subsequent activation of all three PIM kinases. Inhibition of MACC1 and PIM kinase by siRNAs and the effect of IBL-301 (a novel PIM/PI3K/mTOR inhibitor) were examined for effects on cell viability/proliferation and downstream signaling pathways in vitro. Methods: Profiling of H1975P versus H1975GR cells was undertaken using a mRNA/lncRNA array (Arraystar Inc) to quantify the expression of 20,730 mRNAs and 40,173 lncRNAs. Concomitant changes were assessed in the IL6/STAT3 signaling pathway by a QPCR array, immunofluorescence, Western blot and miRNA arrays. The effect of MACC1 and PIM kinase inhibition by siRNAs and targeted agents were examined by Western blot and cell viability assays. Results: MACC1 mRNA was upregulated 77 fold in H1975GR cells after 1-month exposure to apitolisib, and its regulated proteins HGF and MET were upregulated (10.06 and 1.65 fold, respectively). At 4 months MACC1 mRNA was increased by 44 fold, HGF 2 fold and MET 6.6 fold while several members of the IL-6/STAT3 pathway including c-Myc and PIM1, PIM2 and PIM3 kinases were also significantly activated (25.5, 19.8, 3.3 and 31.9 fold respectively). Results were also validated by Western blot. A miRNA signature of PI3K/mTOR inhibitor resistance was validated that can be used to monitor patients on treatment. Inhibition of PIM kinases by siRNA and targeted therapy restored sensitivity to PI3K-mTOR inhibitors. Novel triple-targeted therapy IBL-301 decreased PIM kinase, c-Myc and MACC1 expression. IC50 values of IBL-301 were significantly less for H1975GR cells (0.02μM) compared to H1975P cells (0.75μM), highlighting activated PIM kinase as a mechanism of resistance to PI3K-mTOR inhibition. Conclusion: Activated MACC1, PIM kinases and their interacting partners play a role in resistance to PI3K-mTOR inhibitors in PIK3CA mutant NSCLC. Our data provide a rationale for combined or sequential targeting of PI3K-mTOR and PIM kinases to significantly delay resistance and provide durable treatment responses in patients with PIK3CA mutant lung tumors. Citation Format: Gillian Moore, Samira Elbai, Susan Heavey, Stephen Finn, Michael O'Neill, Kathy Gately. Activation of the MACC1/PIM/cMyc axis confers resistance to PI3K/mTOR inhibition in PIK3CA mutant NSCLC [abstract]. In: Proceedings of the AACR Special Conference on Targeting PI3K/mTOR Signaling; 2018 Nov 30-Dec 8; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(10_Suppl):Abstract nr B14.

Identification of miRNA and mRNA expression profiles by PCR microarray in hepatitis B virus‑associated hepatocellular carcinoma.

The present study aimed to identify differentially expressed microRNAs (miRNAs) and mRNAs in hepatitis B virus‑associated hepatocellular carcinoma (HCC). A total of five HCC tissues and paired adjacent non‑tumor tissues were screened to identify the differentially expressed miRNAs and target mRNAs using polymerase chain reaction microarrays. The interaction between differential miRNA and mRNA expression was concurrently analyzed using bioinformatics methods. A total of 32 differentially expressed miRNAs (four upregulated miRNAs and 28 downregulated miRNAs) and 16 differentially expressed mRNAs (11 upregulated mRNAs and five downregulated mRNAs) were identified. Among these, upregulated hsa‑miRNA (miR)‑96‑5p and hsa‑miR‑18b‑5p suppressed their target mRNAs forkhead box O1 and MET transcriptional regulator MACC1 (MACC1). Downregulation of hsa‑miR‑199a‑5p led to upregulation of its target mRNAs, cyclin dependent kinase 4 and insulin like growth factor 2 (IGF2). The high‑level expression of IGF2 mRNA and cyclin E1 mRNA was due to the low‑level expression of hsa‑miR‑145‑5p, hsa‑miR‑181a‑5p, hsa‑miR‑199a‑5p and hsa‑miR‑223a‑3p, and hsa‑miR‑26a‑5p and hsa‑miR‑26b‑5p, respectively. The low‑level expression of coronin 1A mRNA and MACC1 mRNA was due to overexpression of hsa‑miR‑517a‑3p and hsa‑miR‑18a‑5p, and hsa‑miR‑18b‑5p, respectively. Numerous gene ontology terms were associated with oncogenesis. The most enriched pathways targeted by the dysregulated miRNAs and mRNAs were associated with cancer and oncogenesis pathways. The present data suggested that differential miRNA and mRNA expression is present in HCC. Thus, interactions between certain miRNAs and mRNAs may be involved in the pathogenesis of HCC.

The lncRNA MACC1-AS1 promotes gastric cancer cell metabolic plasticity via AMPK/Lin28 mediated mRNA stability of MACC1

BackgroundMetabolic plasticity has been increasingly thought to be a determinant of tumor growth and metastasis. MACC1, a transcriptional regulator of MET, was recognized as an oncogene in gastric cancer (GC); however, its transcriptional or post-translational regulation was not clear. We previously reported the metabolic role of MACC1 in glycolysis to promote GC progression. MACC1-AS1 is the antisense lncRNA of MACC1, yet its function was previously unknown.MethodsWe profiled and analyzed the expression of MACC1-AS1 utilizing the TCGA database as well as in situ hybridization using 123 pairs of GC tissues and matched adjacent normal gastric mucosa tissues (ANTs). The biological role of MACC1-AS1 in cell growth and metastasis was determined by performing in vitro and in vivo functional experiments. Glycolysis and antioxidant capabilities were assayed to examine its metabolic function. Further, the specific regulatory effect of MACC1-AS1 on MACC1 was explored transcriptionally and post-transcriptionally.ResultsMACC1-AS1 was shown to be expressed significantly higher in GC tissues than in ANTs, which predicted poor prognosis in GC patients. MACC1-AS1 promoted GC cell proliferation and inhibited cell apoptosis under metabolic stress. Mechanistically, MACC1-AS1 stabilized MACC1 mRNA and post-transcriptionally augmented MACC1 expression. Further, MACC1-AS1 was shown to mediate metabolic plasticity through MACC1 upregulation and subsequent enhanced glycolysis and anti-oxidative capabilities, and this was suggested to be coordinated by the AMPK/Lin28 pathway.ConclusionsElevated expression of MACC1-AS1 in gastric cancer tissues is linked to poor prognosis and promotes malignant phenotype upon cancer cells. MACC1-AS1 is elevated under metabolic stress and facilitates metabolic plasticity by promoting MACC1 expression through mRNA stabilization. Our study implicates lncRNA MACC1-AS1 as a valuable biomarker for GC diagnosis and prognosis.

Prognostic value of MACC1 and proficient mismatch repair status for recurrence risk prediction in stage II colon cancer patients: the BIOGRID studies

We assessed the novel MACC1 gene to further stratify stage II colon cancer patients with proficient mismatch repair (pMMR). Four cohorts with 596 patients were analyzed: Charité 1 discovery cohort was assayed for MACC1 mRNA expression and MMR in cryo-preserved tumors. Charité 2 comparison cohort was used to translate MACC1 qRT-PCR analyses to FFPE samples. In the BIOGRID 1 training cohort MACC1 mRNA levels were related to MACC1 protein levels from immunohistochemistry in FFPE sections; also analyzed for MMR. Chemotherapy-naïve pMMR patients were stratified by MACC1 mRNA and protein expression to establish risk groups based on recurrence-free survival (RFS). Risk stratification from BIOGRID 1 was confirmed in the BIOGRID 2 validation cohort. Pooled BIOGRID datasets produced a best effect-size estimate. In BIOGRID 1, using qRT-PCR and immunohistochemistry for MACC1 detection, pMMR/MACC1-low patients had a lower recurrence probability versus pMMR/MACC1-high patients (5-year RFS of 92% and 67% versus 100% and 68%, respectively). In BIOGRID 2, longer RFS was confirmed for pMMR/MACC1-low versus pMMR/MACC1-high patients (5-year RFS of 100% versus 90%, respectively). In the pooled dataset, 6.5% of patients were pMMR/MACC1-low with no disease recurrence, resulting in a 17% higher 5-year RFS [95% confidence interval (CI) (12.6%-21.3%)] versus pMMR/MACC1-high patients (P = 0.037). Outcomes were similar for pMMR/MACC1-low and deficient MMR (dMMR) patients (5-year RFS of 100% and 96%, respectively). MACC1 expression stratifies colon cancer patients with unfavorable pMMR status. Stage II colon cancer patients with pMMR/MACC1-low tumors have a similar favorable prognosis to those with dMMR with potential implications for the role of adjuvant therapy.

Retraction Note to: Evaluation of gene expression level of CDC5L and MACC1 in poor prognosis and progression of osteosarcoma.

Current evidences have indicated that osteosarcoma is strongly associated with abnormal genetic and epigenetic changes that lead to the abnormal expression of oncogenes or methylation of tumor suppressor genes. In the present study, MACC1 and CDC5L mRNA levels in the patients with osteosarcoma were evaluated using quantitative real-time PCR. Our results demonstrated that CDC5L mRNA levels were higher in tumor tissues than in adjacent normal tissues (2.713 ± 0.738 vs. 1.071 ± 0.629; P < 0.05). Moreover, MACC1 was upregulated in tumor bone tissues than in adjacent normal tissues (3.221 ± 0. 624 vs. 1.427 ± 0.456; P < 0.05). Our result demonstrated that high expression of CDC5L was significantly related to advanced TNM stage (P = 0.032). No significant difference was determined between CDC5L mRNA expression and other clinicopathological parameters including age, gender, tumor diameter, location, tumor grade, and histological type. In addition, overexpression of MACC1 was strongly correlated with advanced TNM stage (P = 0.027) and high tumor grade (P = 0.035). Our findings indicated that mRNA level of CDC5L is correlated with advanced TNM stage, and MACC1 may be involved in progression of osteosarcoma.

Effect of colon cancer metastasis associated gene-1 on biological function of gastric cancer cell line MGC-803 by regulating hepatocyte growth factor/c-Met signaling pathway

Objective To study the effect of colon cancer metastasis associated gene-1 (MACC-1) expression on the proliferation and apoptosis of gastric cancer cell line MGC-803 in vitro. Methods Cultured gastric cancer MGC-803 cells and passaged cells were divided into three groups: group A [MACC-1-short hairpin RNA (shRNA) and PSV2neo3 co transfection group]; group B (shNC and PSV2neo co transfection group); group C (non transfection group). By Western blotting and real-time quantitative polymerase chain reaction (Real-time PCR), the expression of MACC-1 protein and mRNA was detected in the three groups, and that of of c-Met, E-cadherin and N-cadherin in 3 groups of MGC-803 cells were detected. Flow cytometry and scratch test were used to analyze migration, proliferation and apotosis of MGC-803 cells transfected with MACC-1-shRNA. Results (1) Western blotting and Real-time PCR test results showed that as compared with groups B and C, MACC-1 and c-Met protein and mRNA expression in group A were significantly down-regulated (P<0.01). (2) Western blotting and Real-time PCR detection results showed as compared with groups B and C, E-cadherin protein and mRNA expression levels were significantly increased (P<0.01), c-Met and N-cadherin mRNA and protein levels were significantly decreased (P<0.01) in group A. There was no significant difference between groups B and C. (3) Scratch assay showed the scratch space was wide, and healing was inhibited in group A. the migration ability of MGC-803 cells in group A was reduced as compared with groups B and C (P<0.01), but there was no statistically significant difference in the scratch space between groups B and C. (4) Flow cytometry revealed that the apoptosis rate in groups A, B and C was (21.06±1.12)%, (2.18±0.14)% and (2.04±0.11%) respectively. The apoptosis rate in group A was significantly higher than in group C (P<0.01), and there was no significant difference between groups B and C. Conclusion The transfection of MACC-1 gene can promote the proliferation and inhibit the apoptosis of gastric cancer cell line MGC-803 cells, which may play a role in the regulation of hepatocyte growth factor (HGF)/c-Met signal transduction pathway. Key words: Gastric cancer; Colon cancer metastasis associated gene-1; Apoptosis; Biological function

Evaluation of gene expression level of CDC5L and MACC1 in poor prognosis and progression of osteosarcoma.

Current evidences have indicated that osteosarcoma is strongly associated with abnormal genetic and epigenetic changes that lead to the abnormal expression of oncogenes or methylation of tumor suppressor genes. In the present study, MACC1 and CDC5L mRNA levels in the patients with osteosarcoma were evaluated using quantitative real-time PCR. Our results demonstrated that CDC5L mRNA levels were higher in tumor tissues than in adjacent normal tissues (2.713 ± 0.738 vs. 1.071 ± 0.629; P < 0.05). Moreover, MACC1 was upregulated in tumor bone tissues than in adjacent normal tissues (3.221 ± 0. 624 vs. 1.427 ± 0.456; P < 0.05). Our result demonstrated that high expression of CDC5L was significantly related to advanced TNM stage (P = 0.032). No significant difference was determined between CDC5L mRNA expression and other clinicopathological parameters including age, gender, tumor diameter, location, tumor grade, and histological type. In addition, overexpression of MACC1 was strongly correlated with advanced TNM stage (P = 0.027) and high tumor grade (P = 0.035). Our findings indicated that mRNA level of CDC5L is correlated with advanced TNM stage, and MACC1 may be involved in progression of osteosarcoma.

Aberrant expression of pim-3 promotes proliferation and migration of ovarian cancer cells.

Pim kinase-3(Pim-3), a member of serine/threonine protein kinases, has been implicated in multiple human cancers and involved in Myc-induced tumorigenesis. However, little is known regarding its expression and biological function in human ovarian cancer. In this study we showed that the clinical significance and biological functions of Pim-3 in ovarian cancer and found that higher Pim-3 mRNA level are detected in ovarian cancer tissues than those in normal ovarian tissues. There are significant correlations between higher Pim-3 expression levels with the FIGO stage, histopathological subtypes, and distant metastasis in ovarian cancer patients. Lentivirus-mediated gene overexpression of Pim-3 significantly promotes the proliferation and migration of SKOV3 cell lines. Furthermore, MACC1 and Pim-3 expression were significantly correlated in human ovarian cancer cells, and overexpression of Pim-3 in ovary cancer cells increased MACC1 mRNA and protein expression. The data indicate that Pim-3 acts as a putative oncogene in ovary cancer and could be a viable diagnostic and therapeutic target for ovarian cancer.

Influence of the MACC1 gene on sensitivity to chemotherapy in human U251 glioblastoma cells.

This study was conducted to determine the influence of MACC1 expression on chemotherapy sensitivity in human U251 glioblastoma cells. Expression of the MACC1 gene in 49 cases of human brain glioma was determined by quantitative real-time PCR. Silencing effects of RNA interference on MACC1 was detected by Western-blotting. Flow cytometry methods and methyl thiazolyl tetrazolium assay (MTT) were used to determine the apoptosis and growth inhibitory rates of the U251 cells with MACC1 silencing. before and after treatment with cisplatin (DDP). MACC1 mRNA in gliomas was up-regulated remarkably, to 158.8% of that in peri-cancerous tissues (P<0.05). The siRNA-MACC1 could inhibit the expression of MACC1 protein significantly (p<0.05), associated with an increase in apoptosis rate from 2.57% to 5.39% in U251 cells and elevation of the growth inhibitory rate from 1.5% to 17.8% (p<0.05 for both). After treatment with DDP at various concentrations (1, 3, 5μg/ml), compared with control U251 cells, the apoptosis rate of MACC1-silenced U251 cells rose from 8.41%, 13.2% and 19.5% to 12.8%, 17.8% and 25.8%; the growth inhibitory rate increased from 16.2%, 19.3% and 24.5% to 23.7%, 28.4% and 36.3%. There is a notable relationship between over-expression of MACC1 and the characteristics of glioma cells. Silencing of MACC1 was found to enhance the apoptosis and growth inhibitory rates of U251 glioma cells, and thereby increase their sensitivity to DDP chemotherapy.

Downregulation of MACC1 expression enhances cisplatin sensitivity in SKOV-3/DDP cells.

The aim of this study was to investigate the correlation between MACC1 expression and resistance to cisplatin (DDP) in DDP-resistant human epithelial ovarian cancer SKOV-3 cells (SKOV-3/DDP). MACC1 mRNA and protein expression levels in SKOV-3 and SKOV-3/DDP cells were detected by reverse transcriptase polymerase chain reaction and western blot. The SKOV-3/DDP cells were divided into 5 groups: control, shVect (transfected with p-super-EGFP-1 plasmid), pshMACC1 (transfected with psuper-EGFP-shMACC1 plasmid), PD (pretreated with 20 μM PD98059), and combined (transfected with psuper-EGFP-shMACC1 plasmid and pretreated with 20 μM PD98059) groups. Cisplatin sensitivity and cell apoptosis in SKOV-3/DDP cells were assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry. ERK1/2 and p-ERK1/2 expression was determined by western blot. MACC1 mRNA and protein expression levels in SKOV-3/DDP cells were 2.66 ± 0.54 and 1.95 ± 0.45 times those seen in SKOV-3 cells (P < 0.05). Cisplatin sensitivity of pshMACC1 group was much higher than that in the control and shVect groups. Cisplatin-induced cell apoptosis rates increased significantly in the pshMACC1, PD, and combined groups, compared to the control and shVect groups. Moreover, the apoptosis rate was the highest in the combined group among the 5 groups (IC50 = 20.836 ± 0.629 μM). p-ERK1/2 expression decreased significantly in the pshMACC1, PD, and combined groups (this decrease was the most obvious in the combined group). In conclusion, downregulation of MACC1 expression could enhance cisplatin sensitivity and decrease drug resistance in SKOV- 3/DDP cells.

Analysis of HGF, MACC1, C-met and apoptosis-related genes in cervical carcinoma mice

To understand the underlying pharmacological basis and the molecular mechanism of Taxol in therapy of cervical carcinoma (CC) disease, we need to explore the effect of Taxol on CC-related genes and pro-apoptosis and anti-apoptosis genes expression. Immunohistochemistry, western blot and reverse transcription-polymerase chain reaction were applied to examine postive expression levels of Bcl-2, Bax and Caspase-3, HGF, MACC1, Caspase-3 and C-met proteins and MACC1 mRNA expression in tumour of CC mice. Results showed that treatment of Taxol could increase the inhibition rate of tumour growth, positive expression levels of Caspase-3, Bax and decrease positive expression levels of Bcl-2 and Bcl-2/Bax, expression levels of HGF, MACC1 and C-met proteins and MACC1 mRNA in tumour tissue of CC mice. It can be concluded that inhibitory activity of Taxol against tumour growth in CC mice is closely associated with its modulating positive expression of Bcl-2, Bax, Caspase-3, expression of HGF, MACC1, Caspase-3 and C-met proteins and MACC1 mRNA in tumour of CC mice. In conclusion, HGF, MACC1 and C-met genes involve into malignant cervical tumors occurrence, development and prognosis, and might become potential molecular target therapy site of cervical cancer. Taxol intervention may serve as a multi-targeted CC therapeutic capable of inducing selective cancer cell death.

MACC1 as a Prognostic Biomarker for Early-Stage and AFP-Normal Hepatocellular Carcinoma

BackgroundThe metastasis-associated in colon cancer 1 gene (MACC1) has been found to be associated with cancer development and progression. The aim of this study was to investigate the prognostic value of MACC1 in early-stage and AFP-normal hepatocellular carcinoma (HCC).MethodsmRNA and protein levels of MACC1 expression in one normal liver epithelial cells THLE3 and 15 HCC cell lines were examined using reverse transcription-PCR and Western blot. MACC1 expression was also comparatively studied in 6 paired HCC lesions and the adjacent non-cancerous tissue samples. Immunohistochemistry was employed to analyze MACC1 expression in 308 clinicopathologically characterized HCC cases. Statistical analyses were applied to derive association between MACC1 expression scores and clinical staging as well as patient survival.ResultsLevels of MACC1 mRNA and protein were higher in HCC cell lines and HCC lesions than in normal liver epithelial cells and the paired adjacent noncancerous tissues. Significant difference in MACC1 expression was found in patients of different TNM stages (P<0.001). Overall survival analysis showed that high MACC1 expression level correlated with lower survival rate (P = 0.001). Importantly, an inverse correlation between MACC1 level and patient survival remained significant in subjects with early-stage HCC or with normal serum AFP level.ConclusionsMACC1 protein may represent a promising biomarker for predicting the prognosis of HCC, including in early-stage and AFP-normal patients.

Abstract 444: MACC1 as biomarker for progression and prognosis of human malignant glioma

Abstract Metastasis-associated in colon cancer-1 (MACC1) is a prognostic indicator for metastasis formation and metastasis-free survival of colon cancer patients. Patients with low MACC1 expressing primary tumors had a 5-year-survival of 80%, whereas subjects with high MACC1 expressing tumors had a 5-year-survival of only 15%. Correlations of MACC1 expression levels with tumor progression, recurrence and metastasis have meanwhile been reported for e.g. gastric, lung and hepatocellular cancer. However, no data are available concerning MACC1 expression in human astrocytic tumors. Glioblastoma multiforme (GBM) are the most prevalent brain tumors of adults either developing de novo without known precursor-lesion (primary GBM), or progressing from low grade astrocytomas (secondary GBM). Patients suffering from GBM have a very disadvantageous prognosis. Although these tumors rarely metastasize, they are characterized by rapid growth, and their invasive and migratory behavior is similar to those of metastatic cells of tumors with different origin. Therefore we tested our hypothesis on MACC1 involvement in progression of human gliomas. MACC1 mRNA expression increased concomitantly with increasing WHO grading, as demonstrated by semi-quantitative and quantitative RT-PCR of biopsies from human low grade astrocytomas and GBM. MACC1 protein expression allowed discrimination of dormant and recurrent LGA and of primary and secondary GBM, as shown by immunohistochemistry of an independent patients’ cohort. Furthermore, strong MACC1 expression was associated with reduced median patients’ survival. Finally, we performed real-time measurements of migration and proliferation in the MACC1-transfected GBM cell line U138/MACC1 and found promoted proliferation and migration. Therefore, MACC1 may be a new biomarker for progression and for prognosis of human malignant gliomas and might represent a new potential therapeutic target for inhibition of their proliferation and migration. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 444. doi:1538-7445.AM2012-444

Overexpression of MACC1 mRNA in lung adenocarcinoma is associated with postoperative recurrence

The purpose of this study was to clarify the role and clinical significance of metastasis associated in colon cancer 1 in resected stage I non-small cell lung cancers. Tumor specimens were collected from 146 consecutive patients who underwent a complete resection for stage I lung adenocarcinoma from 1998 to 2007 at the University of Occupational and Environmental Health. We analyzed the expression of metastasis associated in colon cancer 1 mRNA of primary lung adenocarcinomas by real-time reverse transcriptase-polymerase chain reaction. The average postoperative observation period was 49.4 months. Thirteen (8.9%) of 146 patients had recurrences after surgery. Overexpression of metastasis associated in colon cancer 1 mRNA was identified in 62 patients (42.5%). Metastasis associated in colon cancer 1 was overexpressed in 9 (69.2%) of 13 patients and 53 (39.9%) of 133 patients with and without recurrence, respectively (P = .004). The median metastasis associated in colon cancer 1 copy number was 3.0 and 1.4 in patients with and without tumor recurrence, respectively. Metastasis associated in colon cancer 1 overexpression was associated with poorer disease-free survival according to the survival analysis (P = .033). Metastasis associated in colon cancer 1 gene overexpression may be a useful marker for predicting postoperative recurrence in patients with lung adenocarcinoma after surgery.

Abstract 2258: Targeting MACC1: In vivo imaging of distant metastases reduction by MACC1-shRNA in human colon cancer xenografts

Abstract We identified the previously undescribed gene MACC1 by a genome-wide search for differentially expressed genes in human colon cancer tissues, metastases and normal tissues. MACC1 plays an important role for metastasis prediction, in particular for early-stage patients. Based on MACC1 mRNA expression in not (yet) metastasized primary colon cancers of stages I, II and III, our negative and positive prediction of the development of metachronous distant metastases was correct to 80% and 74%, respectively. The 5-year-survival for subjects suffering from colon cancer was 80% for MACC1 low expressors, but 15% for individuals with high MACC1 expression in their primary tumors. Moreover, we found MACC1 to act as an inducer of migration, invasion and proliferation in cell culture, and of distant metastases in several xenograft models. MACC1 transcriptionally regulates the receptor tyrosine kinase Met, that activates the HGF/Met signaling pathway resulting in enhanced cell motility, invasion and metastasis. Here we report the reduction of metastasis by using shRNA targeting the metastasis-inducer MACC1 in vivo. MACC1 high expressing human colon cancer cells were transfected with MACC1-shRNA, Met-shRNA, or control-shRNA harboring plasmids, together with luciferase-containing constructs. Cell clones were intrasplenically transplanted into mice. Tumor growth and metastases formation was demonstrated by in vivo imaging. Signals in MACC1-shRNA/luc mice and in Met-shRNA/luc mice originated exclusively from spleen tumors, not from liver metastases, as demonstrated with the isolated organs by luminescence. By contrast, signals in control-shRNA/luc mice originated from spleen tumors and liver metastases. These findings were confirmed by brightfield images, demonstrating reduced tumor growth and metastases development in MACC1-shRNA/luc and Met-shRNA/luc mice compared to control-shRNA/luc mice. In summary, knock down of MACC1 by MACC1-shRNA led to reduction of tumor growth and metastasis in mice, making MACC1-shRNA a useful tool for metastasis intervention of colon cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2258.