CCL2 promotes metastasis and epithelial-mesenchymal transition of non-small cell lung cancer via PI3K/Akt/mTOR and autophagy pathways.

In non-small cell lung cancer (NSCLC), metastasis is the most common phenotype, and autophagy plays a vital role in its regulation. However, there are limited data on how autophagy-related genes and metastasis-related genes affect NSCLC progression. Our goal was to identify the genes that regulate autophagy and metastasis in NSCLC, and to assess the underlying mechanisms in this current study. RNA sequencing data from public databases were used to screen differentially expressed autophagy- and metastasis-associated genes. Enrichment analyses and immune correlations were conducted to identify hub genes and potential regulating pathways in NSCLC. In this study, we found that CCL2 expression was highly expressed in NSCLC tissues and high CCL2 level was correlated with strong infiltration in lung tissues from NSCLC patients. Overexpression of CCL2 can enhance the metastasis of NSCLC cells in nude mice. Furthermore, CCL2 activated the PI3K/Akt/mTOR signalling pathway axis, promoted epithelial-mesenchymal transition (EMT), and blocked the autophagic flux in NSCLC cells. Therefore, our results indicate that CCL2 promotes metastasis and EMT of NSCLC via PI3K/Akt/mTOR axis and autophagy signalling pathways. We believe that CCL2 could be a probable target for the diagnosis and therapeutics of NSCLC, and this study may expand our understanding of lung cancer.

Theoretical Insights into the Luminescence and Sensing Mechanisms of N,N'-Bis(salicylidene)-[2-(3',4'-diaminophenyl)benzthiazole] for Copper(II).

The intramolecular proton transfer (IPT) reaction potential energy surfaces (PESs) of N,N'-bis(salicylidene)-[2-(3',4'-diaminophenyl)benzthiazole] (BTS) in the S0 state and S1 state are constructed. It is found that the IPT reactions in the ground state hardly take place due to the high reaction energy barrier for single-proton (6.3 kcal/mol) and double-proton transfer (14.1 kcal/mol) reactions and low backward reaction energy barriers for single-proton (1.9 kcal/mol) and double-proton transfer (1.2 kcal/mol) reactions. In comparison, an excited-state intramolecular single-proton transfer reaction is a barrierless and exothermic process, and thus, single-proton transfer tautomer T1H contributes most to the fluorescence emission. Based on the analysis of PESs, the experimental absorption and emission spectra are reproduced well by the calculated vertical excitation energies of BTS and its photoisomerization products, and the triple fluorescence emission profile in the experiment is reassigned unequivocally. Furthermore, thermodynamic analysis of the BTS-Cu(II) complex shows that the dinuclear complex (C1) with Cu(II) coordinating with O and N atoms of the hydrogen bonds is the most thermodynamically stable structure, and the intramolecular hydrogen bonding structure in BTS is destroyed due to the chelation of Cu(II) and BTS; as a result, the IPT reaction of C1 in S0 and S1 states is significantly inhibited. The inhibitor of Cu(II) in the IPT reaction plays a major role in fluorescence quenching.

Microfluidic platform for studying the anti-cancer effect of ursolic acid on tumor spheroid.

At present, the probability that a new anti-tumor drug will eventually succeed in clinical trials is extremely low. In order to make up for this shortcoming, the use of a three-dimensional (3D) cell culture model for secondary screening is often necessary. Cell spheroid is the easiest 3D model tool for drug screening. In this study, the microfluidic chip with a microwell array was manufactured, which could allow the formation of tumor spheroids with uniform size and easily retrieve cell spheroids from the chip. Cell spheroids were successfully cultured for over 15 days and the survival rate was as high as 80%. Subsequently, cellular response to the ursolic acid (UA) was observed on the chip. Compared to the monolayer culture cells in vitro, the tumor spheroids showed minor levels of epithelial-mesenchymal transition fluctuation after drug treatment. The mechanism of cell spheroid resistance to UA was further verified by detecting the expression level of upstream pathway proteins. But the invasive ability of tumor spheroids was attenuated when the duration of action of UA extended. The anti-cancer effect of UA was innovatively evaluated on breast cancer by using the microfluidic device, which could provide a basis and direction for future preclinical research on UA.

Quantitative Prediction of Charge Mobilities and Theoretical Insight into the Regulation of Site-Specific Trifluoromethylethynyl Substitution to Electronic and Charge Transport Properties of 9,10-Anthraquinone.

Herein, we systematically studied the electronic and conducting properties of 9,10-anthraquinone (AQ) and its derivatives and discussed the substitute-site effects on their organic field-effect transistor (OFET) properties in detail. Our calculation results show the influence of different substitute sites on the ionization potential (IP), electronic affinity (EA), reorganization energy (λ), electronic couplings (V), and anisotropic mobility (µ) of semiconducting materials, which mainly originates from the variations of the frontier molecular orbital charge distributions, the steric hindrance, and the conjugate degree. Combining quantum-chemical calculations with charge transfer theory, we simulated the intermolecular hopping rate in the organic crystals of AQ derivatives and predicted the fluctuation range of three-dimensional (3D) anisotropic charge carrier mobility for the first time. Our calculation results well reproduced the experimental observations and provided evidence for the determination of the optimal OFET conduction plane and channel direction relative to the crystal axis.

Theoretical study of charge-transport and optical properties of organic crystals: 4,5,9,10-pyrenedi-imides.

This work presents a systematic study of the conducting and optical properties of a family of aromatic di-imides reported recently and discusses the influences of side-chain substitution on the reorganization energies, crystal packing, electronic couplings and charge injection barrier of 4,5,9,10-pyrenedi-imide (PyDI). Quantum-chemical calculations combined with the Marcus-Hush electron transfer theory revealed that the introduction of a side chain into 4,5,9,10-pyrenedi-imide increases intermolecular steric interactions and hinders close intermolecular π-π stacking, which results in weak electronic couplings and finally causes lower intrinsic hole and electron mobility in t-C5-PyDI (µh = 0.004 cm2 V-1 s-1 and µe = 0.00003 cm2 V-1 s-1) than in the C5-PyDI crystal (µh = 0.16 cm2 V-1 s-1 and µe = 0.08 cm2 V-1 s-1). Furthermore, electronic spectra of C5-PyDI were simulated and time-dependent density functional theory calculation results showed that the predicted fluorescence maximum of t-C5-PyDI, corresponding to an S 1→S 0 transition process, is located at 485 nm, which is close to the experimental value (480 nm).

DNA fragment-assisted microfluidic chip for capture and release of circulating tumor cells.

Circulating tumor cells are specifically referred as cells that detached from the primary tumor and are present in the bloodstream. They could be isolated from blood and used as representative biomarker for predicting cancer prognoses. Here, we developed a microfluidic chip with multiple curved channels, in which DNA fragments and antibody-based enrichment are exploited to capture circulating tumor cells in blood sample. By introducing DNA fragments as long tentacles, the active antibody could be extended into the microchannel stereoscopically, which could greatly increase the chances of adhesion in a multidirectional way and improve the capture efficacy. Several pivotal factors for cell capturing were optimized to the best state. Compared to conventional chips for planar capturing, the capture efficiency of MCF-7 cells was greatly increased from 37.17 to 85.10%. For the detection of MCF-7-containing artificial blood sample detection, the capture efficiency of tumor cells was about 74.19 ± 2.13%, which was obviously better than the result of flow cytometry (29.67 ± 4.02%). Captured cells were easily released from the surface of microfluidic chip with high cell viability, which could be investigated for the molecular analysis in the field of tumor diagnosis.

Electrical-field tuned thermoelectric performance of graphene nanoribbon with sawtooth edges.

By using first-principle calculations combined with the non-equilibrium Green's function approach, we report that a vertical electrical field can modulate the thermoelectric performance of a graphene nanoribbon with sawtooth edges. The results show that the sawtooth graphene nanoribbon exhibits the spin-dependent Seebeck effect under the temperature gradients, and is strengthened by increasing the width of the sawtooth graphene nanoribbon. When the vertical electrical field is applied to the device, the spin-dependent Seebeck effect can also be enhanced. The vertical electrical field can modulate the device transferring from hole-conducting to electron-conducting. This opens up the possibility of tuning the thermal transport properties of the device by the electrical field.

Facile synthesis of fluorescent carbon dots from Prunus cerasifera fruits for fluorescent ink, Fe3+ ion detection and cell imaging.

Carbon dots (CDs) synthesized from natural products have drawn numerous attentions due to some unique properties. Here, Prunus cerasifera fruits were used as carbon source to synthesize high luminescent CDs by hydrothermal method. The obtained CDs were characterized by TEM, FTIR and XPS methods, founding the CDs were near-spherical and contained abundant nitrogen element. The CDs aqueous solution exhibited bright blue fluorescence under ultraviolet illumination, with the maximum emission at 450 nm. They could be potentially used as invisible fluorescent ink by written on the paper and irradiated by UV light, due to their fluorescent properties. Moreover, the CDs were found being selectively quenched by Fe3+ ion. The quench of CDs was linearly related to the concentration of Fe3+ ion in the range of 0-0.5 mM, meaning they could be developed as fluorescent probe of Fe3+ ion. At last, the CDs were used for cell imaging, founding they were low toxicity to HepG2 cells and exhibited blue and green fluorescence under a fluorescence microscope. In summary, the CDs prepared from Prunus cerasifera fruits exhibited excellent fluorescence properties, and could be potentially applied in the field of fluorescent ink, Fe3+ ion detection and cell imaging.

In situ monitoring of the structural change of microemulsions in simulated gastrointestinal conditions by SAXS and FRET.

Microemulsions are promising drug delivery systems for the oral administration of poorly water-soluble drugs. However, the evolution of microemulsions in the gastrointestinal tract is still poorly characterized, especially the structural change of microemulsions under the effect of lipase and mucus. To better understand the fate of microemulsions in the gastrointestinal tract, we applied small-angle X-ray scattering (SAXS) and fluorescence resonance energy transfer (FRET) to monitor the structural change of microemulsions under the effect of lipolysis and mucus. First, the effect of lipolysis on microemulsions was studied by SAXS, which found the generation of liquid crystalline phases. Meanwhile, FRET spectra indicated micelles with smaller particle sizes were generated during lipolysis, which could be affected by CaCl2, bile salts and lecithin. Then, the effect of mucus on the structural change of lipolysed microemulsions was studied. The results of SAXS and FRET indicated that the liquid crystalline phases disappeared, and more micelles were generated. In summary, we studied the structural change of microemulsions in simulated gastrointestinal conditions by SAXS and FRET, and successfully monitored the appearance and disappearance of the liquid crystalline phases and micelles.

Long noncoding RNA HOTAIR promotes renal cell carcinoma malignancy through alpha-2, 8-sialyltransferase 4 by sponging microRNA-124.

OBJECTIVES: Accumulating evidence demonstrated that the long noncoding RNA (lncRNA) HOTAIR (Hox transcript antisense intergenic RNA) plays key role in renal cell carcinoma (RCC) malignancy, while microRNA-124 (miR-124) is a tumour suppressor in RCC. The aim of this work was to assess the biological function of HOTAIR and to explore underlying mechanism involved in HOTAIR/miR-124/alpha-2, 8-sialyltransferase 4 (ST8SIA4) axis-regulated progression in RCC. MATERIALS AND METHODS: Real-time PCR analyses and western blots were performed to the levels of HOTAIR, miR-124 and ST8SIA4 expression in human RCC tissues and RCC cell lines (ACHN and 786-O). Bioinformatics analysis and dual-luciferase reporter assay were used to illustrate relationship between HOTAIR and miR-124 in RCC. Colony formation assays, EdU assays, Ki67 assays and apoptosis assays were taken to evaluate cell proliferation. Tumour xenograft was created to explore the functions of HOTAIR and ST8SIA4 in tumorigenesis in vivo. Migration assays, invasion assays and cell adhesion assays and were also taken to analyse the carcinoma progression. RESULTS: In this study, HOTAIR level was confirmed to be significantly upregulated in RCC samples and RCC cell lines compared with those in the paired adjacent tissues and normal renal cell line. Overexpression of HOTAIR promoted the capability of proliferation, migration and invasion in RCC cell lines. HOTAIR directly bound to miR-124, while miR-124 mediated the expression of ST8SIA4 in RCC cell lines. ST8SIA4 was upregulated in RCC tissues and RCC cell lines. Ectopic expression of ST8SIA4 modulated the proliferation, migration and invasion of RCC cells. Further results indicated that HOTAIR promoted the proliferation and metastasis as a competing endogenous RNA to regulate ST8SIA4 expression by sponging miR-124 in RCC. CONCLUSIONS: Our results demonstrated that HOTAIR mediated RCC progression in part through miR-124/ST8SIA4 axis, which functioned as a new prognostic biomarker in RCC.

A microfluidic chip-based co-culture of fibroblast-like synoviocytes with osteoblasts and osteoclasts to test bone erosion and drug evaluation.

Targeting fibroblast-like synoviocyte (FLS) migration and invasion-mediated bone erosion is a promising clinical strategy for the treatment of rheumatoid arthritis (RA). Drug sensitivity testing is fundamental to this scheme. We designed a microfluidic chip-based, cell co-cultured platform to mimic RA FLS-mediated bone erosion and perform drug-sensitive assay. Human synovium SW982 cells were cultured in the central channel and migrated to flow through matrigel-coated side channels towards cell culture chamber where RANKL-stimulated osteoclastic RAW264.7 and osteogenic medium (OS)-stimulated bone marrow mesenchymal stem cells (BMSC) were cultured in the microfluidic chip device, mimicking FLS migration and invasion-mediated bone erosion in RA. These SW982 cells showed different migration potentials to osteoclasts and BMSC. The migration of SW982 cells with high expression of cadherin-11 was more potent when SW982 cells were connected with the co-culture of RAW264.7 and BMSC. Simultaneously, in the co-cultured chamber, tartrate-resistant acid phosphatase (TRAP) activity of RANKL-stimulated RAW264.7 cells was enhanced, but alkaline phosphatase (ALP) activity was decreased in comparison with mono-cultured chamber. Furthermore, it was confirmed that celastrol, a positive drug for the treatment of RA, inhibited SW982 cell migration as well as TRAP activity in the cell-cultured microfluidic chips. Thus, the migration and invasion to bone-related cells was reconstituted on the microfluidic model. It may provide an effective anti-RA drug screen model for targeting FLS migration-mediated bone erosion.

Charge-transport properties of 4-(1,2,2-tri-phenyl-vinyl)-aniline salicylaldehyde hydrazone: tight-packing induced molecular 'hardening'.

Based on first-principles calculations, the relationship between molecular packing and charge-transport parameters has been investigated and analysed in detail. It is found that the crystal packing forces in the flexible organic molecule 4-(1,2,2-triphenylvinyl)-aniline salicylaldehyde hydrazone (A) can apparently overcome the dynamic intramolecular rotations and the intramolecular steric repulsion, effectively enhancing the molecular rigidity and decreasing the internal reorganization energy. The conducting properties of A have also been simulated within the framework of hopping models, and the calculation results show that the intrinsic electron mobility in A is much higher than the corresponding intrinsic hole mobility. These theoretical investigations provide guidance for the efficient and targeted control of the molecular packing and charge-transport properties of organic small-molecule semiconductors and conjugated polymeric materials.

Preparation of Essential Oil-Based Microemulsions for Improving the Solubility, pH Stability, Photostability, and Skin Permeation of Quercetin.

Quercetin can bring many benefits to skin based on its various bioactivities. However, the therapeutic effect of quercetin is limited due to the poor water solubility, pH instability, light instability, and skin permeation. The aim of the present work was applying essential oil-based microemulsions to improve the solubility, pH stability, photostability, and skin permeation of quercetin for topical application. Peppermint oil (PO-ME), clove oil (CO-ME), and rosemary oil (RMO-ME) were selected as model essential oils. Microemulsions composed of Cremophor EL/1,2-propanediol/essential oils (47:23:30, w/w) were selected as model formulations, based on the pseudo-ternary phase diagram and the characterizations. In the solubility study, the solubility of quercetin was improved dozens of times by microemulsions. Quercetin was found instable under alkaline condition, with 50% degraded in the solution of pH 13. However, PO-ME, CO-ME, and RMO-ME could protect quercetin from the hydroxide ions, with 47, 9, and 12% of quercetin degraded. In the photostability study, the essential oil-based microemulsions showed the capability of protecting quercetin from degradation under UV radiation. Where more than 67% of quercetin was degraded in aqueous solution, while less than 7% of quercetin degraded in microemulsions. At last, the in vitro skin permeation study showed that the essential oil-based microemulsions could enhance the permeation capacity of quercetin by 2.5-3 times compared to the aqueous solution. Hence, the prepared essential oil microemulsions could improve the solubility, pH stability, photostability, and skin permeation of quercetin, which will be beneficial for its topical application.

A DFT Study on the Electronic Structures and Conducting Properties of Rubrene and its Derivatives in Organic Field-Effect Transistors.

We systematically studied the electronic structures and conducting properties of rubrene and its derivatives reported recently, and disscussed the influences of electron-withdrawing groups and chemical oxidation on the reorganization energies, crystal packing, electronic couplings, and charge injection barrier of rubrene. Hirshfeld surface analysis and quantum-chemical calculations revealed that the introduction of CF3 groups into rubrene decreases the H···H repulsive interaction and increases intermolecular F···H/H···F attractive interactions, which resulted in the tight packing arrangement and the increase of the electronic couplings, and finally cause the higer intrinsic hole-mobility in bis(trifluoromethyl)-dimethyl-rubrene crystal (µh = 19.2 cm2 V-1 s-1) than in rubrene crystal (µh = 15.8 cm2 V-1 s-1). In comparison, chemical oxidation reduces charge-carrier mobility of rubrene crystal by 2~4 orders of magnitude and increased the hole and electron injection barrier, which partly explains the rubrene-based field-effect transistor performance degrades upon exposure to air. Furthermore, we also discussed the influence of structural parameters of carbon nanotube (CNT) electrode on charge injection process, which suggests that the regulation of CNT diameters and increasing in thickness is an effective strategy to optimize CNT work functions and improve n-type OFET performances based on these organic materials.

Wedelolactone Enhances Osteoblastogenesis but Inhibits Osteoclastogenesis through Sema3A/NRP1/PlexinA1 Pathway.

Bone remodeling balance is maintained by tight coupling of osteoblast-mediated bone formation and osteoclast-mediated bone resorption. Thus, agents with the capacity to regulate osteoblastogenesis and osteoclastogenesis have been investigated for therapy of bone-related diseases such as osteoporosis. In this study, we found that wedelolactone, a compound isolated from Ecliptae herba, and a 9-day incubation fraction of conditioned media obtained from wedelolactone-treated bone marrow mesenchymal stem cell (BMSC) significantly inhibited tartrate-resistant acid phosphatase (TRAP) activity in RANKL-stimulated osteoclastic RAW264.7 cells. Addition of the semaphorin 3A (Sema3A) antibody to the conditioned media partially blocked the medium's inhibitory effects on the RAW264.7 cells. In BMSC, mRNA expression of Sema3A increased in the presence of different wedelolactone concentrations. Blocking Sema3A activity with its antibody reversed wedelolactone-induced alkaline phosphatase activity in BMSC and concurrently enhanced wedelolactone-reduced TRAP activity in osteoclastic RAW264.7 cells. Moreover, in BMSC, wedelolactone enhanced binding of Sema3A with cell-surface receptors, including neuropilin (NRP)1 and plexinA1. Furthermore, nuclear accumulation of ß-catenin, a transcription factor acting downstream of wedelolactone-induced Sema3A signaling, was blocked by the Sema3A antibody. In osteoclastic RAW264.7 cells, conditioned media and wedelolactone promoted the formation of plexin A1-NRP1, but conditioned media also caused the sequestration of the plexin A1-DNAX-activating protein 12 (DAP12) complex and suppressed the phosphorylation of phospholipase C (PLC)γ2. These data suggest that wedelolactone promoted osteoblastogenesis through production of Sema3A, thus inducing the formation of a Sema3A-plexinA1-Nrp1 complex and ß-catenin activation. In osteoclastic RAW264.7 cells, wedelolactone inhibited osteoclastogenesis through sequestration of the plexinA1-DAP12 complex, induced the formation of plexinA1-Nrp1 complex, and suppressed PLCγ2 activation.

Induction of epithelial-to-mesenchymal transition in proximal tubular epithelial cells on microfluidic devices.

In proteinuric nephropathy, epithelial-to-mesenchymal transition (EMT) is an important mechanism that causes renal interstitial fibrosis. The precise role of EMT in the pathogenesis of fibrosis remains controversial, partly due to the absence of suitable in vitro or in vivo models. We developed two microfluidic and compartmental chips that reproduced the fluidic and three-dimensional microenvironment of proximal tubular epithelial cells in vivo. Using one microfluidic device, we stimulated epithelial cells with a flow of healthy human serum, heat-inactivated serum and complement C3a, which mimicked the flow of urine within the proximal tubule. We observed that epithelial cells exposed to serum proteins became apoptotic or developed a mesenchymal phenotype. Incubating cells with C3a induced similar features. However, cells exposed to heat-inactivated serum did not adopt the mesenchymal phenotype. Furthermore, we successfully recorded the cellular morphological changes and the process of transmigration into basement membrane extract during EMT in real-time using another three-dimensional microdevice. In conclusion, we have established a cell-culture system that mimics the native microenvironment of the proximal tubule to a certain extent. Our data indicates that EMT did occur in epithelial cells that were exposed to serum proteins, and C3a plays an essential role in this pathological process.

A simple microfluidic strategy for cell migration assay in an in vitro wound-healing model.

In vitro scratch wound assays are commonly used strategies to measure cell repair rate, facilitating the study of cell migration, tissue reorganization, and cell division. This work presented a simple and novel microfluidic device that allowed a quantitative investigation of the cell migration and cell proliferation behaviors in an in vitro wound-healing model, especially focused on the scratch assay. The microfluidic device is composed of four units, which include cell growth regions and cell-free regions created by micropillars. Using this device, we evaluated the proliferation and migration process of human gastric epithelial cells in the presence of different concentrations of the epidermal growth factor, and investigated the migration behavior of mesenchymal stem cells toward tumor cells as well. This approach has the unique capability to create localized cell-free regions in parallel, and facilitate quantitative research on cell migration in the wound-healing process, providing a powerful platform for elucidating the mechanism of cell migration in regeneration medicine.

Biomimetic tumor microenvironment on a microfluidic platform.

Tumor microenvironment is a highly complex system consisting of non-cancerous cells, soluble factors, signaling molecules, extracellular matrix, and mechanical cues, which provides tumor cells with integrated biochemical and biophysical cues. It has been recognized as a significant regulator in cancer initiation, progression, metastasis, and drug resistance, which is becoming a crucial component of cancer biology. Modeling microenvironmental conditions of such complexity in vitro are particularly difficult and technically challenging. Significant advances in microfluidic technologies have offered an unprecedented opportunity to closely mimic the physiological microenvironment that is normally encountered by cancer cells in vivo. This review highlights the recent advances of microfluidic platform in recapitulating many aspects of tumor microenvironment from biochemical and biophysical regulations. The major events relevant in tumorigenesis, angiogenesis, and spread of cancer cells dependent on specific combinations of cell types and soluble factors present in microenvironmental niche are summarized. The questions and challenges that lie ahead if this field is expected to transform the future cancer research are addressed as well.

Probing the role of mesenchymal stem cells in salivary gland cancer on biomimetic microdevices.

Mesenchymal stem cells (MSCs) are the progenitors of stromal cells, which have been found to interact with cancer cells and represent an important target for cancer therapies. Salivary gland cancer is an aggressive malignant epithelial tumor and can easily disseminate to distant sites in vivo. In this work, we probed the role of MSCs in salivary gland cancer in an in vivo like tumor microenvironment by using a series of functional microfluidic devices for 2D and 3D assays. The results demonstrated that MSCs could be recruited by salivary gland cancer cells (ACC-M) and this effect was potentially mediated by TGF-ß secreted by cancer cells. In addition, MSCs exhibited the ability to squeeze into ACC-M spheroids by dispersing the cell-cell connection and reducing the expression of E-cadherin in cancer cells. In particular, MSCs exhibited the ability to enhance the invasion of salivary cancer under a chemokine CXCL12 gradient, indicating the involvement of a CXCL12-CXCR4 pathway and the tumor-promoting properties of MSCs in cancer progression. This study recreated an in vivo-like tumor microenvironment by constructing a series of functional microdevices and explored the role of MSCs in ACC invasion for the first time. It provides a unique platform to open up new options to target stromal cells for cancer therapy and also analyzed the potential risk of using MSCs as drug delivery carriers for therapeutic purposes in carcinoma treatment.

Characterization of the interaction between fibroblasts and tumor cells on a microfluidic co-culture device.

Fibroblasts and tumor cells have been involved in the process of cancer development, progression and therapy. Here, we present a simple microfluidic device which enables to study the interaction between fibroblasts and tumor cells by indirect contact co-culture. The device is composed of multiple cell culture chambers which are connected by a parallel of cell migration regions, and it enables to realize different types of cells to communicate each other on the single device. In this work, human embryonic lung fibroblasts cells were observed to exhibit obvious migration towards tumor cells instead of normal epithelial cells on the co-culture device. Moreover, transdifferentiation of human embryonic lung fibroblast cells was recognized by the specific expression of alpha-smooth muscle actin, indicating the effect of tumor cells on the behavior of fibroblasts. Furthermore, multiple types of cell co-culture can be demonstrated on the single device which enables to mimic the complicated microenviroment in vivo. The device is simple and easy to operate, which enables to realize real-time observation of cell migration after external stimulus. This microfluidic device allows for the characterization of various cellular events on a single device sequentially, facilitating the better understanding of interaction between heterotypic cells in a more complex microenvironment.