Estimation of cancer cell migration in biomimetic random/oriented collagen fiber microenvironments

Increasing data indicate that cancer cell migration is regulated by extracellular matrixes and their surrounding biochemical microenvironment,playing a crucial role in pathological processes such as tumor invasion and metastasis.However,conventional two-dimensional cell culture and animal models have limitations in studying the influence of tumor microenvironment on cancer cell migration.Fortunately,the further development of microfluidic technology has provided solutions for the study of such questions.We utilize microfluidic chip to build a random collagen fiber microenvironment(RFM)model and an oriented collagen fiber microenvironment(OFM)model that resemble early stage and late stage breast cancer microenvironments,respectively.By combining cell culture,biochemical concentration gradient construction,and microscopic imaging techniques,we investigate the impact of different collagen fiber biochemical microenvironments on the migration of breast cancer MDA-MB-231-RFP cells.The results show that MDA-MB-231-RFP cells migrate further in the OFM model compared to the RFM model,with significant differences observed.Furthermore,we establish concentration gradients of the anticancer drug paclitaxel in both the RFM and OFM models and find that paclitaxel significantly inhibits the migration of MDA-MB-231-RFP cells in the RFM model,with stronger inhibition on the high concentration side compared to the low concentration side.However,the inhibitory effect of paclitaxel on the migration of MDA-MB-231-RFP cells in the OFM model is weak.These findings suggest that the oriented collagen fiber microenvironment resembling the late-stage tumor microenvironment is more favorable for cancer cell migration and that the effectiveness of anticancer drugs is diminished.The RFM and OFM models constructed in this study not only provide a platform for studying the mechanism of cancer development,but also serve as a tool for the initial measurement of drug screening.

The saponin DT-13 inhibits gastric cancer cell migration through down-regulation of CCR5-CCL5 axis

AIM: To investigate the effect of DT-13 on gastric cancer cell migration, and to explore the possible mechanisms underlying the anti-metastasis activity of DT-13. METHODS: Growth inhibition of DT-13 was analyzed by the MTT assay. Cell migration was measured by the scratch-wound assay and transwell double chamber assay. To investigate the possible mechanisms underlying the anti-metastasis activity of DT-13, chemokine receptors that are involved in cancer metastasis(CCR2, CCR5, CCR7, CXCR4, and CXCR6) were detected by conventional PCR. The effect of DT-13 on CCR5 and CXCR4 expression was further evaluated by quantitative PCR and Western blot, respectively. The secretion of CCL5(ligand of CCR5) and SDF-1(ligand of CXCR4) were detected by enzyme-linked immunosorbent assay(ELISA). RESULTS: DT-13 inhibited BGC-823 and HGC-27 cell growth in a dose dependent manner, and the estimated IC50 value for 24 h treatment was 23.5 ± 5.1 μmol·L-1 for BGC-823 cells and 35.6 ± 7.6 μmol·L-1 for HGC-27 cells. DT-13 also significantly decreased gastric cancer cell migration. DT-13 significantly decreased the gene expression of CCR5 in both BGC-823 and HGC-27 gastric cancer cells, and moderately reduced the expression of CXCR4. Similar to the results of gene expression, significant down-regulation of CCR5 protein was observed, but CXCR4 protein levels were much less affected. CCL5 secretion, but not SDF-1 production, was inhibited by DT-13. CONCLUSION: DT-13 inhibited gastric cancer cell migration by down-regulation of the CCR5-CCL5 axis.

Mechanical transmission enables EMT cancer cells to drive epithelial cancer cell migration to guide tumor spheroid disaggregation

Cell phenotype heterogeneity within tumor tissue,especially which due to the emergence of epithelial-mesenchymal transition(EMT) in cancer cells,is associated with cancer invasion and metastasis.However,our understanding of the cellular mechanism(s)underlying the cooperation between EMTcell and epithelial cancer cell migration remains incomplete.Herein,heterotypic tumor spheroids containing both epithelial and EMT cancer cells were generated in vitro.We observed that EMT cells dominated the peripheral region of the self-organized heterotypic tumor spheroid.Furthermore,our results demonstrated that EMT cells could serve as leader cells to improve the collective migration efficiency of epithelial cancer cells and promote dispersion and invasion of the tumor spheroids,which was regulated by the force transition between EMT cells and epithelial cancer cells.Mechanistically,our data further suggest that force transmission is mediated by heterophilic N-cadherin/E-cadherin adhesion complexes between EMT and epithelial cancer cells.Impairment of N-cadherin/E-cadherin adhesion complex formation abrogated the ability of EMT cells to guide epithelial cancer cell migration and blocked the dispersion of tumor spheroids.Together,our data provide new insight into the mechanical interaction between epithelial and EMT cancer cells through heterophilic cadherin adhesion,which enables cooperative tumor cell migration,highlighting the role of EMT cells in tumor invasion.

Knockdown of GRHL3 Inhibits Activities and Induces Cell Cycle Arrest and Apoptosis of Human Colorectal Cancer Cells

The Grainyhead-like 3（GRHL3） is involved in epidermal barrier formation, neural tube closure and wound repair. Previous studies have suggested that GRHL3 has been linked to many different types of cancers. However, to date, its effects on human colorectal cancer（CRC） has not been clarified yet. Our microarray analysis has indicated predominant GRHL3 expression in CRC. The purpose of this study was to investigate the expression and significance of GRHL3 in CRC tumorigenesis using CRC tissues and paired paracancerous tissues, as well as using distinct CRC cell lines（HT29 and DLD1）. We observed increased GRHL3 expression at both m RNA and protein levels in CRC tissues and CRC cell lines using quantitative real-time polymerase chain reaction（q RT-PCR） and Western blotting. Moreover, silencing GRHL3 with si RNA could suppress CRC cell proliferation, viability and migration in vitro. We also found that knockdown of GRHL3 could promote cell cycle arrest at G0/G1 phase in HT29 cells and DLD1 cells, and induce cell apoptosis in HT29 cells. Together, our study revealed the down-regulation of GRHL3 in vitro could inhibit CRC cell activity and trigger cell cycle arrest at G0/G1 phase and apoptosis.

CD74 and macrophage migration inhibitory factor as therapeutic targets in gastric cancer

AIM:To investigate the relationship and molecular features of CD74/macrophage migration inhibitory factor(MIF)/Toll-like receptor 4(TLR4) in gastric cancer.METHODS:CD74,MIF and TLR4 expression in the paraffin-embedded sections of gastric cancer from 120 patients were detected by immunohistochemical staining.Knock down of CD74 expression in gastric cancer cell line MKN-45 was performed by lentivirus transduction and detected by Western blotting.MKN-45 cell proliferation assay under the stimulants was measured by the cell counting kit 8(CCK8) assay and MIF concentration in the culture medium was detected by enzymelinked immunosorbent assay.Surface staining of CD74 in the MKN-45 cell line under the stimulation of lipopolysaccharide(LPS) was measured by flow cytometry.MIF,CD74 and TLR4 co-localization in the MKN-45 cell line was performed by the immunoprecipitation.RESULTS:CD74,MIF and TLR4 were found to be expressed in gastric cancer and increased significantly in the advanced stage,and were also associated with lymph node metastasis.Correlation analysis revealed that CD74 was positively correlated with MIF(r = 0.2367,P < 0.01) and both proteins were also associated with TLR4(r = 0.4414,r = 0.5001,respectively,P < 0.01).LPS can significantly promote MKN-45 cell proliferation(3.027 ± 0.388 vs 4.201 ± 0.092,P < 0.05),induce MIF production(54.333 ± 2.906 pg/mL vs 29.667 ± 3.180 pg/mL,P < 0.01) and cell surface expression of CD74(75.6% ± 4.046%vs 9.4% ± 0.964%,P < 0.01) at LPS concentration of 1 μg/mL compared to medium control.Knockdown of CD74 or using antiCD74 and MIF antagonist ISO-1 significantly reduced LPS-induced MKN-45 cell proliferation(4.201 ± 0.092 vs 3.337 ± 0.087,4.534 ± 0.222 vs 3.368 ± 0.290,4.058 ± 0.292vs 2.934 ± 0.197,respectively,P < 0.01).MIF,CD74 and TLR4 could co-localize in the MKN-45 cell line.CONCLUSION:Upregulation of MIF,CD74 and TLR4 are associated with increasing clinical stage and provide an opportunity as novel gastric cancer chemoprevention and/or treatment strategy.

Effects of bisphenol compounds on the growth and epithelial mesenchymal transition of MCF-7 CV human breast cancer cells

Bisphenol-A（BPA） has been considered as an endocrine disrupting chemical（EDC） because it can exert estrogenic properties.For bisphenol-S（BPS） and bisphenol-F（BPF） that are BPA analogs and substitutes,their risk to estrogendependent cancer has been reported rarely compared with the numerous cases of BPA.In this study,we examined whether BPA,BPS,and BPF can lead to the proliferation,migration,and epithelial mesenchymal transition（EMT） of MCF-7 clonal variant（MCF-7 CV） breast cancer cells expressing estrogen receptors（ERs）.In a cell viability assay,BPA,BPS,and BPF significantly increased proliferation of MCF-7 CV cells compared to control（DMSO） as did17β-estradiol（E2）.In Western blotting assay,BPA,BPS,and BPF enhanced the protein expression of cell cycle progression genes such as cyclin D1 and E1.In addition,MCF-7 CV cells lost cell to cell contacts and acquired fibroblast-like morphology by the treatment of BPA,BPS,or BPF for 24 hours.In cell migration assay,BPA,BPS,and BPF accelerated the migration capability of MCF-7 CV cells as did E2.In relation with the EMT process,BPA,BPS,and BPF increased the protein expression of N-cadherin,while they decreased the protein expression of Ecadherin.When BPA,BPS,and BPF were co-treated with ICI 182,780,an ER antagonist,proliferation effects were reversed,the expression of cyclin D1 and cyclin E1 was downregulated,and the altered cell migration and expression of N-cadherin and E-cadherin by BPA,BPS,and BPF were restored to the control level.Thus,these results imply that BPS and BPF also have the risk of breast cancer progression as much as BPA in the induction of proliferation and migration of MCF-7 CV cells by regulating the protein expression of cell cycle-related genes and EMT markers via the ER-dependent pathway.

The role of YAP in the control of the metastatic potential of oral cancer

The Yes-associated protein(YAP)is a downstream effector of the Hippo pathway and acts as a key transcription co-factor to regulate cell migration,proliferation,and survival.The Hippo pathway is evolutionarily conserved and controls tissue growth and organ size.Dysregulation and heterogeneity of this pathway are found in cancers,including oral squamous cell carcinoma(OSCC),leading to overexpression of YAP and its regulated proliferation machinery.The activity of YAP is associated with its nuclear expression and is negatively regulated by the Hippo kinase-mediated phosphorylation resulting in an induction of its cytoplasmic translocation.This review focuses on the role of YAP in OSCC in the context of cancer metastatic potential and highlights the latestfindings about the heterogeneity of YAP expression and its nuclear transcription activity in oral cancer cell lines.The review also discusses the potential target of YAP in oral cancer therapy and the recentfinding of the unprecedented role of the desmosomal cadherin desmoglein-3(DSG3)in regulating Hippo-YAP signaling.

Effects of human microRNA-181a-5p on proliferation and migration of gastric cancer cells

Objective To preliminarily explore the effects of human microRNA-181a on migration of gastric cancer cells and its mechanism.Methods The expression of miRNA-181a-5p in gastric cancer cell line GC9811 and peritoneal high metastasis gastric cancer cell line GC9811-P were tested by quantitative real-time polymerase chain reaction(qRT-PCR).GC9811 cell line was

A review of microfluidic approaches for investigating cancer extravasation during metastasis

Metastases,or migration of cancers,are common and severe cancer complications.Although the 5-year survival rates of primary tumors have greatly improved,those of metastasis remain below 30%,highlighting the importance of investigating specific mechanisms and therapeutic approaches for metastasis.Microfluidic devices have emerged as a powerful platform for drug target identification and drug response screening and allow incorporation of complex interactions in the metastatic microenvironment as well as manipulation of individual factors.In this work,we review microfluidic devices that have been developed to study cancer cell migration and extravasation in response to mechanical(section‘Microfluidic investigation of mechanical factors in cancer cell migration’),biochemical(section‘Microfluidic investigation of biochemical signals in cancer cell invasion’),and cellular(section‘Microfluidic metastasis-on-a-chip models for investigation of cancer extravasation’)signals.We highlight the device characteristics,discuss the discoveries enabled by these devices,and offer perspectives on future directions for microfluidic investigations of cancer metastasis,with the ultimate aim of identifying the essential factors for a‘metastasis-on-a-chip’platform to pursue more efficacious treatment approaches for cancer metastasis.