Divergent behaviors and underlying mechanisms of cell migration and invasion in non-metastatic T24 and its metastatic derivative T24T bladder cancer cell lines

Abstract

Previous studies on cancer cell invasion were primarily focused on its migration because these two events were often considered biologically equivalent. Here we found that T24T cells exhibited higher invasion but lower migration abilities than T24 cells. Expression of Rho-GDPases was much lower and expression of SOD2 was much higher in T24T cells than those in T24 cells. Indeed, knockdown of SOD2 in T24T cells can reverse the cell migration but without affecting cell invasion. We also found that SOD2 inhibited the JNK/c-Jun cascade, and the inhibition of c-Jun activation by ectopic expression of TAM67 impaired Rho-GDPases expression and cell migration in T24T shSOD2 cells. Further, we found that Sp1 can upregulate SOD2 transcription in T24T cells. Importantly, matrix metalloproteinase-2 (MMP-2) was overexpressed in T24T and participated in increasing its invasion, and MMP-2 overexpression was mediated by increasing nuclear transport of nucleolin, which enhanced mmp-2 mRNA stability. Taken together, our study unravels an inverse relationship between cell migration and invasion in human bladder cancer T24T cells and suggests a novel mechanism underlying the divergent roles of SOD2 and MMP-2 in regulating metastatic behaviors of human bladder T24T in cell migration and invasion.

Isorhapontigenin (ISO) Inhibits Invasive Bladder Cancer Formation In Vivo and Human Bladder Cancer Invasion In Vitro by Targeting STAT1/FOXO1 Axis

“…Elevated MMP-2 levels have been reported to be of independent prognostic value in patients with BCs (33), while the other essential MMP member, such as MMP9 expression, is not correlated with tumor grade, stage, or overall survival of the patients with BC (34). Our previous study has also confirmed that up-regulation of MMP-2, but not MMP9, participated in mediation of BC invasion (22). To determine whether MMP-2 is associated with ISO inhibition of human BC invasion, we evaluated the effect of ISO on MMP-2 protein expression.…”

“…The human metastatic BC cell line T24T, which was a lineage-related lung metastatic variant of invasive BC cell line T24 (20-22), was kindly provided by Dr. Dan Theodorescu (Departments of Urology, University of Virginia, Charlottesville, VA) (20) in 2010. All the cell lines were subjected to DNA tests and authenticated in our previous studies (4).…”

Induction of miR-137 by Isorhapontigenin (ISO) Directly Targets Sp1 Protein Translation and Mediates Its Anticancer Activity BothIn VitroandIn Vivo

“…T24T was kindly provided by Dr. Dan Theodorescu (Department of Urology, University of Virginia, Charlottesville, VA, USA) in 2010 and used in our previous studies (4, 5, 23). The UMUC3 cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS), 2mM L-glutamine, and 25μg/ml of gentamycin and the T24T cell was cultured in a 1:1 mixture of Dulbecco’s modified Eagle’s medium (DMEM)/Ham’s F12 medium supplemented with 5% FBS, 2mM L-glutamine, and 25 μg/ml of gentamycin.…”

“…The cell extracts were subjected to SDS-polyacrylamide gels (SDS-PAGE), transferred to polyvinylidene fluoride (PVDF) membranes (Bio-Rad, Hercules, CA, USA), probed with the indicated primary antibodies, and incubated with the AP-conjugated secondary antibody. The protein band specifically bound to the primary antibody was detected by Typhoon FLA 7000 (GE Healthcare, Pittsburgh, PA, USA) using an alkaline phosphatase-linked secondary antibody and an enhanced chemifluorescence Western blotting system as described in our previous studies (5, 23). …”

Inhibition of PHLPP2/cyclin D1 protein translation contributes to the tumor suppressive effect of NFκB2 (p100)

“…Primary p100+/+ and p100−/− mouse embryonic fibroblasts (MEFs) were immortalized according to the modified 3T3-immortalized protocol [46]. Human bladder cancer cell line UMUC3 and T24 cells were described in our previous studies [47, 48]. All cell lines were authenticated 4 to 6 months before use by using a short tandem repeat method.…”