MiR-215-5p Reduces Liver Metastasis in an Experimental Model of Colorectal Cancer through Regulation of ECM-Receptor Interactions and Focal Adhesion

Background: Growing evidence suggests that miR-215-5p is a tumor suppressor in colorectal cancer (CRC); however, its role in metastasis remains unclear. This study evaluates the effects of miR-215 overexpression on the metastatic potential of CRC. Methods: CRC cell lines were stably transfected with miR-215-5p and used for in vitro and in vivo functional analyses. Next-generation sequencing and RT-qPCR were performed to study changes on the mRNA level. Results: Overexpression of miR-215-5p significantly reduced the clonogenic potential, migration, and invasiveness of CRC cells in vitro and tumor weight and volume, and liver metastasis in vivo. Transcriptome analysis revealed mRNAs regulated by miR-215-5p and RT-qPCR confirmed results for seven selected genes. Significantly elevated levels of CTNNBIP1 were also observed in patients’ primary tumors and liver metastases compared to adjacent tissues, indicating its direct regulation by miR-215-5p. Gene Ontology and KEGG pathway analysis identified cellular processes and pathways associated with miR-215-5p deregulation. Conclusions: MiR-215-5p suppresses the metastatic potential of CRC cells through the regulation of divergent molecular pathways, including extracellular-matrix-receptor interaction and focal adhesion. Although the specific targets of miR-215-5p contributing to the formation of distant metastases must be further elucidated, this miRNA could serve as a promising target for CRC patients’ future therapeutic strategies.

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LncRNA MIR17HG promotes colorectal cancer liver metastasis by mediating a glycolysis-associated positive feedback circuit

Oncogene ◽

10.1038/s41388-021-01859-6 ◽

2021 ◽

Author(s):

Senlin Zhao ◽

Bingjie Guan ◽

Yushuai Mi ◽

Debing Shi ◽

Ping Wei ◽

...

Keyword(s):

Colorectal Cancer ◽

Liver Metastasis ◽

Positive Feedback ◽

Feedback Loop ◽

Metastatic Tumor ◽

Feedback Circuit ◽

Colorectal Cancer Liver Metastasis ◽

Positive Feedback Circuit

AbstractGlycolysis plays a crucial role in reprogramming the metastatic tumor microenvironment. A series of lncRNAs have been identified to function as oncogenic molecules by regulating glycolysis. However, the roles of glycolysis-related lncRNAs in regulating colorectal cancer liver metastasis (CRLM) remain poorly understood. In the present study, the expression of the glycolysis-related lncRNA MIR17HG gradually increased from adjacent normal to CRC to the paired liver metastatic tissues, and high MIR17HG expression predicted poor survival, especially in patients with liver metastasis. Functionally, MIR17HG promoted glycolysis in CRC cells and enhanced their invasion and liver metastasis in vitro and in vivo. Mechanistically, MIR17HG functioned as a ceRNA to regulate HK1 expression by sponging miR-138-5p, resulting in glycolysis in CRC cells and leading to their invasion and liver metastasis. More interestingly, lactate accumulated via glycolysis activated the p38/Elk-1 signaling pathway to promote the transcriptional expression of MIR17HG in CRC cells, forming a positive feedback loop, which eventually resulted in persistent glycolysis and the invasion and liver metastasis of CRC cells. In conclusion, the present study indicates that the lactate-responsive lncRNA MIR17HG, acting as a ceRNA, promotes CRLM through a glycolysis-mediated positive feedback circuit and might be a novel biomarker and therapeutic target for CRLM.

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ROS/PI3K/Akt and Wnt/β-catenin signalings activate HIF-1α-induced metabolic reprogramming to impart 5-fluorouracil resistance in colorectal cancer

Journal of Experimental & Clinical Cancer Research ◽

10.1186/s13046-021-02229-6 ◽

2022 ◽

Vol 41 (1) ◽

Author(s):

Shuohui Dong ◽

Shuo Liang ◽

Zhiqiang Cheng ◽

Xiang Zhang ◽

Li Luo ◽

...

Keyword(s):

Colorectal Cancer ◽

Glucose Metabolism ◽

Pentose Phosphate ◽

Metabolic Reprogramming ◽

Cell Models ◽

Primary Tumors ◽

Pharmacological Inhibition ◽

Potential Biomarker

Abstract Background Acquired resistance of 5-fluorouracil (5-FU) remains a clinical challenge in colorectal cancer (CRC), and efforts to develop targeted agents to reduce resistance have not yielded success. Metabolic reprogramming is a key cancer hallmark and confers several tumor phenotypes including chemoresistance. Glucose metabolic reprogramming events of 5-FU resistance in CRC has not been evaluated, and whether abnormal glucose metabolism could impart 5-FU resistance in CRC is also poorly defined. Methods Three separate acquired 5-FU resistance CRC cell line models were generated, and glucose metabolism was assessed by measuring glucose and lactate utilization, RNA and protein expressions of glucose metabolism-related enzymes and changes of intermediate metabolites of glucose metabolite pool. The protein levels of hypoxia inducible factor 1α (HIF-1α) in primary tumors and circulating tumor cells of CRC patients were detected by immunohistochemistry and immunofluorescence. Stable HIF1A knockdown in cell models was established with a lentiviral system. The influence of both HIF1A gene knockdown and pharmacological inhibition on 5-FU resistance in CRC was evaluated in cell models in vivo and in vitro. Results The abnormality of glucose metabolism in 5-FU-resistant CRC were described in detail. The enhanced glycolysis and pentose phosphate pathway in CRC were associated with increased HIF-1α expression. HIF-1α-induced glucose metabolic reprogramming imparted 5-FU resistance in CRC. HIF-1α showed enhanced expression in 5-FU-resistant CRC cell lines and clinical specimens, and increased HIF-1α levels were associated with failure of fluorouracil analog-based chemotherapy in CRC patients and poor survival. Upregulation of HIF-1α in 5-FU-resistant CRC occurred through non-oxygen-dependent mechanisms of reactive oxygen species-mediated activation of PI3K/Akt signaling and aberrant activation of β-catenin in the nucleus. Both HIF-1α gene knock-down and pharmacological inhibition restored the sensitivity of CRC to 5-FU. Conclusions HIF-1α is a potential biomarker for 5-FU-resistant CRC, and targeting HIF-1a in combination with 5-FU may represent an effective therapeutic strategy in 5-FU-resistant CRC.

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Elevated expression of AGGF1 predicts poor prognosis and promotes the metastasis of colorectal cancer

10.21203/rs.2.14319/v4 ◽

2019 ◽

Author(s):

Xin Zhang ◽

Huimin Sun ◽

Wanyuan Chen ◽

Xianglei He

Keyword(s):

Colorectal Cancer ◽

Distant Metastasis ◽

Mrna Level ◽

Disease Free Survival ◽

Normal Mucosa ◽

Expression Level ◽

Angiogenic Factor ◽

Migration And Invasion

Abstract Background: Angiogenic factor with G-patch and FHA domains 1 (AGGF1) can promote angiogenesis and increasing evidence has highlighted the important roles of AGGF1 in tumorigenesis. However, the differential expression as well as the biological functions of AGGF1 in colorectal cancer (CRC) remain to be established. The purpose of the present study is therefore to identify the effect of AGGF1 on prognosis and metastasis in CRC patients. Methods: The expression level of AGGF1 in CRC was examined by qPCR, western blot and immunohistochemistry in a tissue microarray containing 236 CRC specimens and paired normal mucosae. And the effect of AGGF1 on CRC cell malignance was investigated in our established stable AGGF1 upregulated and knockdown CRC cell lines. Results: The expression level of AGGF1 in CRC tissue was not significantly different to that in adjacent normal mucosa at the mRNA level. However, at the protein level, AGGF1 expression in CRC tissues was significantly higher than in paired normal mucosa, which showed a clear association with TNM stage, AJCC stage, vascular invasion, and differentiation. Further, we revealed an apparent correlation between AGGF1 expression and poorer disease-free survival and overall survival of CRC patients. In addition, we discovered that AGGF1 significantly promoted CRC cell wound healing, migration, and invasion in vitro and distant metastasis in vivo. Conclusions: Our study demonstrates the aberrant overexpression of AGGF1 in CRC and provides a basis on which to explore the application of AGGF1 as a potential therapeutic target for CRC patients, especially for CRC patients with distant metastasis.

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LINC01605, regulated by the EP300-SMYD2 complex, potentiates the binding between METTL3 and SPTBN2 in colorectal cancer

Cancer Cell International ◽

10.1186/s12935-021-02180-8 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Meng Yue ◽

Tao Liu ◽

Guoqiang Yan ◽

Xiaofan Luo ◽

Lei Wang

Keyword(s):

Colorectal Cancer ◽

Metastatic Potential ◽

Rna Seq ◽

Related Factors ◽

Non Coding Rna ◽

Long Non Coding Rna ◽

Tumor Related Death ◽

Validation Experiments

Abstract Background Colorectal cancer (CC) is one of the major contributors to tumor-related death worldwide, and its main cause of death is distant metastasis. Dysregulation of long non-coding RNA (lncRNA) LINC01605 has been implicated in CC. However, its role in metastasis of CC remains elusive. The goal of the study is to uncover the biological function and molecular mechanism of LINC01605 in CC. Methods The differentially expressed lncRNAs were first screened from GSE97300, GSE84983, GSE110715, GSE70880, and GSE75970 microarrays. The correlation between the expression of LINC01605 and the clinical phenotypes of enrolled CC patients (n = 134) was subsequently analyzed. The upstream and downstream regulatory mechanisms of LINC01605 in CC were identified through bioinformatics and RNA-seq analyses. Finally, the effects of related factors on CC cell growth and metastasis were confirmed through functional validation experiments. Results LINC01605, significantly highly expressed in CC, was a prognostic factor for patients with CC. Functional experiments revealed that LINC01605 knockdown inhibited the proliferatory and metastatic potential of CC cells in vitro and in vivo. Moreover, LINC01605 was regulated by SMYD2-EP300-mediated modifications of histone H3K4me3 as well as H3K27ac. LINC01605 was found to bind to METTL3 and promote the m6A modification of SPTBN2 mRNA, thereby facilitating the translation of SPTBN2. Conclusions Overexpression of LINC01605, regulated by SMYD2-EP300-mediated H3K27ac and H3K4me3 modifications, bound to METTL3 protein to promote m6A modification of SPTBN2 mRNA, leading to the development of CC.

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Hepatic Scaling Factors for In Vitro-In Vivo Extrapolation (IVIVE) of Metabolic Drug Clearance in Patients with Colorectal Cancer with Liver Metastasis

Drug Metabolism and Disposition ◽

10.1124/dmd.121.000359 ◽

2021 ◽

pp. DMD-AR-2021-000359

Author(s):

Areti-Maria Vasilogianni ◽

Brahim Achour ◽

Daniel Scotcher ◽

Sheila Annie Peters ◽

Zubida M. Al-Majdoub ◽

...

Keyword(s):

Colorectal Cancer ◽

Liver Metastasis ◽

Drug Clearance ◽

Scaling Factors ◽

Metabolic Drug ◽

In Vivo Extrapolation

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A new KSRP-binding compound suppresses distant metastasis of colorectal cancer by targeting the oncogenic KITENIN complex

Molecular Cancer ◽

10.1186/s12943-021-01368-w ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Jeong A Bae ◽

Woo Kyun Bae ◽

Sung Jin Kim ◽

Yoo-Seung Ko ◽

Keon Young Kim ◽

...

Keyword(s):

Colorectal Cancer ◽

Liver Metastasis ◽

Distant Metastasis ◽

Regulatory Protein ◽

Binding Pocket ◽

In Vitro Assays ◽

Docking Model ◽

Small Molecule Compound

Abstract Background Distant metastasis is the major cause of death in patients with colorectal cancer (CRC). Previously, we identified KITENIN as a metastasis-enhancing gene and suggested that the oncogenic KITENIN complex is involved in metastatic dissemination of KITENIN-overexpressing CRC cells. Here, we attempted to find substances targeting the KITENIN complex and test their ability to suppress distant metastasis of CRC. Methods We screened a small-molecule compound library to find candidate substances suppressing the KITENIN complex in CRC cells. We selected a candidate compound and examined its effects on the KITENIN complex and distant metastasis through in vitro assays, a molecular docking model, and in vivo tumor models. Results Among several compounds, we identified DKC1125 (Disintegrator of KITENIN Complex #1125) as the best candidate. DKC1125 specifically suppressed KITENIN gain of function. After binding KH-type splicing regulatory protein (KSRP), DKC1125 degraded KITENIN and Dvl2 by recruiting RACK1 and miRNA-124, leading to the disintegration of the functional KITENIN–KSRP–RACK1–Dvl2 complex. A computer docking model suggested that DKC1125 specifically interacted with the binding pocket of the fourth KH-domain of KSRP. KITENIN-overexpressing CRC cells deregulated certain microRNAs and were resistant to 5-fluorouracil, oxaliplatin, and cetuximab. DKC1125 restored sensitivity to these drugs by normalizing expression of the deregulated microRNAs, including miRNA-124. DKC1125 effectively suppressed colorectal liver metastasis in a mouse model. Interestingly, the combination of DKC1125 with 5-fluorouracil suppressed metastasis more effectively than either drug alone. Conclusion DKC1125 targets the KITENIN complex and could therefore be used as a novel therapeutic to suppress liver metastasis in CRC expressing high levels of KITENIN.

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Prognostic Value and Biological Function of LRRN4 in Colorectal Cancer

10.21203/rs.3.rs-1010837/v1 ◽

2021 ◽

Author(s):

Cheng Xu ◽

Yulin Chen ◽

Feiwu Long ◽

Junman Ye ◽

Xue Li ◽

...

Keyword(s):

Colorectal Cancer ◽

Mrna Level ◽

Biological Significance ◽

Progression Free Survival ◽

Molecular Determinant ◽

Chinese Cohort ◽

Xenograft Models

Abstract Background: Several nervous and nerve-related biomarkers have been detected in colorectal cancer (CRC) and can contribute to the progression of CRC. However, the role of leucine-rich repeat neuronal 4 (LRRN4), a recently identified neurogenic marker, in CRC remains unclear. Methods: We examined the expression and the clinical outcomes of LRRN4 in CRC from TCGA-COREAD mRNA-sequencing datasets and immunohistochemistry on the Chinese cohort. Furthermore, the colony formation, flow cytometry, wound healing assay and mouse xenograft models were used to investigate the biological significance of LRRN4 in CRC cell lines with LRRN4 knockdown or overexpression in vitro and in vivo. In addition, weighted co-expression network analysis and DAVID were used to explore the potential molecular mechanism. Results: We provide the first evidence that LRRN4 expression, at both the protein and mRNA level, was remarkable high in CRC compared to controls and positively correlated with the clinical outcome of CRC patients. Specifically, LRRN4 was an independent prognostic factor for progression-free survival and overall survival in CRC patients. Further functional experiments showed that LRRN4 promoted cell proliferation, cell DNA synthesis and cell migration and inhibited apoptosis. Knockdown of LRRN4 can correspondingly decrease these effects in vitro and can significantly suppress the growth of xenografts. Several biological functions and signaling pathways were regulated by LRRN4, including proteoglycans in cancer, glutamatergic synapse, Ras, MAPK and PI3K.Conclusions: Our results suggest that LRRN4 could be a biological and molecular determinant to stratify CRC patients into distinct risk categories, and mechanistically, this is likely attributable to LRRN4 in regulating several malignant phenotypes of neoplastic cells via cancer-related pathways.

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Micro-PET imaging of angiogenesis based on 18F-RGD for assessment liver metastasis in colorectal cancer

10.21203/rs.3.rs-29007/v1 ◽

2020 ◽

Author(s):

Mingyu Zhang ◽

Hao Jiang ◽

Huijie Jiang ◽

Rongjun Zhang ◽

Zhenchang Wang

Keyword(s):

Colorectal Cancer ◽

Liver Metastasis ◽

Tumor Imaging ◽

Pearson Correlation ◽

Positron Emission Tomography Imaging ◽

Metastatic Potential ◽

Mice Model ◽

Primary Tumors ◽

Ki67 Expression ◽

Standardized Uptake Values

Abstract Background: This study aimed to explore the feasibility of 18 F-AIF-NOTA-E[PEG4-c(RGDfk)]2 (denoted as 18 F-RGD) PET quantitative parameters to distinguish the angiogenesis in colorectal cancer (CRC) mice which has different metastatic potential. Methods: Animal models of CRC liver metastases were established by implanttation of human CRC cell lines LoVo and LS174T via intrasplenic injection. Radiotracer-based micro-positron emission tomography imaging of animal model was performed and the uptake of 18 F-RGD tracer in the tumor tissues were quantified as tumor-to-liver maximum or mean standardized uptake values ratio. Pearson correlation was used to analyze the relationship between radioactive parameters and tumor markers. Results: The SUVmax ratio and SUVmean ratio of LoVo model was significantly higher than LS174T ones in both liver metastasis and primary tumor lesions (P ＜ 0.05 ). A significantly difference was observed in both VEGF and Ki67 expression between LoVo and LS174T primary tumors (P ＜ 0.05 ). The T/L SUVmean or SUVmax ratio of 18 F-RGD showed a significantly correlation with VEGF expression, but weakly correlated with Ki67 expression. The areas under the ROC curves of 18 F-RGD SUVmean ratio for differentiate LoVo from LS174T tumor was 0.801. Conclusions: The T/L SUVmean ratio of 18 F-RGD is a promising parameters for tumor imaging and monitoring angiogenesis process in CRC xenograft mice model.

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The Role and Therapeutic Potential of miRNAs in Colorectal Liver Metastasis

Scientific Reports ◽

10.1038/s41598-019-52225-2 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 3

Author(s):

Smiti S. Sahu ◽

Shatovisha Dey ◽

Sarah C. Nabinger ◽

Guanglong Jiang ◽

Alison Bates ◽

...

Keyword(s):

Colorectal Cancer ◽

Liver Metastasis ◽

Cell Lines ◽

Therapeutic Potential ◽

Metastatic Potential ◽

Sequencing Project ◽

Cancer Genome Sequencing ◽

Novel Therapeutic Strategies

Abstract Colorectal cancer (CRC) is the fourth leading cause of cancer-related deaths worldwide. Liver metastasis is the major cause of CRC patient mortality, occurring in 60% patients with no effective therapies. Although studies have indicated the role of miRNAs in CRC, an in-depth miRNA expression analysis is essential to identify clinically relevant miRNAs and understand their potential in targeting liver metastasis. Here we analyzed miRNA expressions in 405 patient tumors from publicly available colorectal cancer genome sequencing project database. Our analyses showed miR-132, miR-378f, miR-605 and miR-1976 to be the most significantly downregulated miRNAs in primary and CRC liver metastatic tissues, and CRC cell lines. Observations in CRC cell lines indicated that ectopic expressions of miR-378f, -605 and -1976 suppress CRC cell proliferation, anchorage independent growth, metastatic potential, and enhance apoptosis. Consistently, CRC patients with higher miR-378f and miR-1976 levels exhibited better survival. Together, our data suggests an anti-tumorigenic role of these miRNAs in CRC and warrant future in vivo evaluation of the molecules for developing biomarkers or novel therapeutic strategies.

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Elevated expression of AGGF1 predicts poor prognosis and promotes the metastasis of colorectal cancer

BMC Cancer ◽

10.1186/s12885-019-6474-7 ◽

2019 ◽

Vol 19 (1) ◽

Author(s):

Xin Zhang ◽

Huimin Sun ◽

Wanyuan Chen ◽

Xianglei He

Keyword(s):

Colorectal Cancer ◽

Distant Metastasis ◽

Mrna Level ◽

Disease Free Survival ◽

Normal Mucosa ◽

Expression Level ◽

Angiogenic Factor ◽

Migration And Invasion

Abstract Background Angiogenic factor with G-patch and FHA domains 1 (AGGF1) can promote angiogenesis and increasing evidence has highlighted the important roles of AGGF1 in tumorigenesis. However, the differential expression as well as the biological functions of AGGF1 in colorectal cancer (CRC) remain to be established. The purpose of the present study is therefore to identify the effect of AGGF1 on prognosis and metastasis in CRC patients. Methods The expression level of AGGF1 in CRC was examined by qPCR, western blot and immunohistochemistry in a tissue microarray containing 236 CRC specimens and paired normal mucosae. And the effect of AGGF1 on CRC cell malignance was investigated in our established stable AGGF1 upregulated and knockdown CRC cell lines. Results The expression level of AGGF1 in CRC tissue was not significantly different to that in adjacent normal mucosa at the mRNA level. However, at the protein level, AGGF1 expression in CRC tissues was significantly higher than in paired normal mucosa, which showed a clear association with TNM stage, AJCC stage, vascular invasion, and differentiation. Further, we revealed an apparent correlation between AGGF1 expression and poorer disease-free survival and overall survival of CRC patients. In addition, we discovered that AGGF1 significantly promoted CRC cell wound healing, migration, and invasion in vitro and distant metastasis in vivo. Conclusions Our study demonstrates the aberrant overexpression of AGGF1 in CRC and provides a basis on which to explore the application of AGGF1 as a potential therapeutic target for CRC patients, especially for CRC patients with distant metastasis.

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