Validation of the effects of TGF-β1 on tumor recurrence and prognosis through tumor retrieval and cell mechanical properties

Abstract

BackgroundIn vivo, the transforming growth factor-beta1 (TGF-β1)-induced epithelial to mesenchymal transition (EMT) occurs in seconds during cancer cells intravasation and extravasation. Although it has been established that cellular stiffness can change as a cancer cell transformed, the precise relationship between TGF-β1-induced mesenchymal stem cell mechanics and cancer prognosis remains unclear. Accordingly, it is hard to define the effects of EMT on cell mechanical properties (CMs), tumor recurrence and metastasis risks. This study bridges physical and pathological disciplines to reconcile single-cell mechanical measurements of tumor cells.Methods and resultsWe developed a microplate measurement system (MMS) and revealed the intrinsic divergent tumor composition of retrieval cells by cell stiffness and adhesion force and flow cytometry analysis. After flow cytometry sorting, we could measure the differences in CMs of the Sca-1+-CD44+ (mesenchymal-stem-cell-type) and the other subgroups. As well as the stiffer and heterogeneous compositions among tumor tissues with higher recurrence risk were depicted by MMS and atomic force microscopy (AFM). An in vitro experiment validated that Lewis lung carcinoma (LLC) cells acquired higher CMs and motility after EMT, but abrogated by SB-505124 inhibition. Concomitantly, the CD31, MMP13 and TGF-β1 enriched micro-environment in the tumor was associated with higher recurrence and distal lung metastasis risks. Furthermore, we report a comprehensive effort to correlate CMs to tumor-prognosis indicators, in which a decreased body weight gain ratio (BWG) and increased tumor weight (TW) were correlated with increased CMs.ConclusionsTogether, we determined that TGF-β1 was significantly associated with malignant tumor progressing. In terms of clinical applications, local tumor excision followed by MMS analysis offers an opportunity to predict tumor recurrence and metastasis risks.

The relationship between vasculogenic mimicry and epithelial‐mesenchymal transitions

“…It has been revealed that doxycycline inhibits EMT‐related transcription factor activity and that doxycycline exerts its antitumour effect by interfering with tumour cell EMT. These findings show that doxycycline acts upstream of EMT‐related signal transduction to inhibit a wide range of cellular functions . In addition, methacycline appear to have similar inhibitory effects on TGF‐β1‐induced EMT in both A549 cells and lung epithelial cells compared with doxycycline, although doxycycline is not as potent as methacycline .…”

“…These findings show that doxycycline acts upstream of EMT-related signal transduction to inhibit a wide range of cellular functions [81][82][83]. In addition, methacycline appear to have similar inhibitory effects on TGF-b1-induced EMT in both A549 cells and lung epithelial cells compared with doxycycline, although doxycycline is not as potent as methacycline [84].…”

LncRNA LINC00472 regulates cell stiffness and inhibits the migration and invasion of lung adenocarcinoma by binding to YBX1

“…YBX1 also activates the translation of Snail mRNA to directly induce EMT 31 . Previous studies showed that the cell stiffness changed during the EMT process 15 . To investigate whether the interaction between LINC00472 and YBX1 affected the regulation of Snail, we performed qRT-PCR, western blot and immunofluorescence assays.…”

“…These changes may affect the surface topography and mechanical properties of the cells. Previous studies showed that the stiffness of the cells changes when the cells undergo EMT 15 . TGF-β is a key factor in the induction of EMT in tumor cells 41 .…”

“…LncRNAs have attracted attention for their new role in cancer 11 , 12 . Increasing studies showed that lncRNAs are involved in cancer progress via regulation of the occurrence of the EMT 13 , 14 , which is related to the biophysical characteristics of cells 15 .…”