Sulforaphane metabolites inhibit migration and invasion via microtubule-mediated Claudins dysfunction or inhibition of autolysosome formation in human non-small cell lung cancer cells

Abstract

Both sulforaphane-cysteine (SFN-Cys) and sulforaphane-N-acetyl-L-cysteine (SFN-NAC) inhibited cancer migration and invasion, but the underlying mechanisms were not clear. Here we uncovered via tissue microarray assay that high expression of invasion-associated Claudin-5 was correlated to malignant grades in human non-small cell lung cancer (NSCLC) cells. Further, SFN-Cys (10 µM) induced the accumulated phosphorylation of ERK1/2, leading to downregulation of Claudin-5 and upregulation of Claudin-7, and the decrease of Claudin-1 in SK-1 cells and increase of Claudin-1 in A549 cells; knockdown of Claudin-5 significantly reduced invasion, whereas knockdown of Claudin-7 increased invasion; knockdown of Claudin-1 reduced invasion in SK-1 cells, whereas it increased invasion in A549 cells, indicating that SFN-Cys regulated Claudins and inhibited invasion depending on Claudin isotypes and cell types. Furthermore, immunofluorescence staining showed that SFN-Cys triggered microtubule disruption and knockdown of α-tubulin downregulated Claudin-1, 5, and 7, and inhibited migration and invasion, indicating that microtubule disruption contributed to invasive inhibition. Co-immunoprecipitation and confocal microscopy observation showed that SFN-Cys lowered the interaction between α-tubulin and Claudin-1 or 5, or 7. Meanwhile, Western blotting and immunofluorescence staining showed that SFN-NAC (15 µM) downregulated α-tubulin resulting in microtubule disruption; knockdown of α-tubulin increased SFN-NAC-induced LC3 II accumulation in SK-1 cells. Combined with the inhibitor of autolysosome formation, Bafilomycin A1 (100 nM), SFN-NAC inhibited invasion via accumulating LC3 II and blocking formation of autolysosome. Further, SFN-NAC upregulated microtubule-stabilizing protein Tau; knockdown of Tau reduced LC3 II/LC3 I inhibiting migration and invasion. These results indicated that SFN-Cys inhibited invasion via microtubule-mediated Claudins dysfunction, but SFN-NAC inhibited invasion via microtubule-mediated inhibition of autolysosome formation in human NSCLC cells.

Sulforaphane-cysteine inhibited migration and invasion via enhancing mitophagosome fusion to lysosome in human glioblastoma cells

“…Recently, we reported that SFN metabolites induced apoptosis and inhibited migration and invasion via causing microtubule disruption 7,13,25 ; here we further determined that SFN-Cys inhibited the invasion in GBM by phosphorylated ERK1/2-mediated downregulation of Stathmin-1 and α-tubulin, and microtubule disruption. Stathmin-1 stimulated microtubule catastrophes leading to promotion of microtubule disassembly, while pStathmin-1 (Ser 25) blocked the role of Stathmin-1 34 .…”

“…Claudin-5 was determined to interact with the MT1 matrix metalloproteinase increasing cell migration and invasion via degradation of the extracellular matrix 16,17 . We found that knockdown of α-tubulin downregulated Claudins and inhibited migration and invasion, indicating that microtubule disruption contributed to invasive inhibition in NSCLC cells 7 . The cytosolic C-terminal domain of Claudin-5 contains a PDZ-binding domain which binds ZO-1, ZO-2, and ZO-3 proteins, consequently binding to the microtubules 18 .…”

“…Even, autophagy promotes metastatic tumor recurrence; knockdown of autophagy regulators inhibits GBM migration and invasion 24 . We previously revealed that SFN metabolites inhibited autophagy leading to migration and invasion 7 . Autophagy promotes focal adhesion disassembly and cell motility of metastatic tumor cells through the direct interaction of Paxillin to LC3 37 .…”

MicroRNAs as novel targets of sulforaphane in cancer therapy: The beginning of a new tale?

“…In terms of antitumor activity, sulforaphane has shown great potential for treatment of cancers such as bladder cancer (Xia, Kang, Jung, Zhang, & Lian, ), colon cancer (Darkwa, Burkhardt, & Tsuji, ), triple negative breast cancer (Jensen, Slattery, Housley, & Hansen, ), pancreatic cancer (Yin et al, ), prostate cancer (Traka et al, ), and non‐small cell lung cancer (Zheng et al, ). Although high numbers of studies have investigated the antitumor activity of sulforaphane, there is a need for a precise review to describe the mechanism of antitumor activity of sulforaphane.…”

Integrated oncological treatment in the course of metastatic mammary neoplasia: a complete resolution case

“…In recent years, it has progressively emerging that the combined use of clinically-relevant anticancer treatments in association with natural substances, acupuncture techniques and adequate nutrition, has led to the resolution of cases with metastatic diffusion. Substances, such as turmeric (Curcuma longa), graviola (Annona muricata), Boswellia serrata, and sulforaphane have been reported to have a potential antitumoral activity [2][3][4][5][6][7]. Several studies are focused on the 'integrative oncology' a term that has coined to identify the use of complementary and integrative therapies in combination with conventional oncological care [8,9].…”