Sublethal doxorubicin promotes migration and invasion of breast cancer cells: role of Src Family non-receptor tyrosine kinases

Abstract

Background Doxorubicin (Dox) is a widely used chemotherapy, but its effectiveness is limited by dose-dependent side effects. Although lower Dox doses reduce this risk, studies have reported higher recurrence of local disease with no improvement in survival rate in patients receiving low doses of Dox. To effectively mitigate this, a better understanding of the adverse effects of suboptimal Dox doses is needed. Methods Effects of sublethal dose of Dox on phenotypic changes were assessed with light and confocal microscopy. Migratory and invasive behavior were assessed by wound healing and transwell migration assays. MTT and LDH release assays were used to analyze cell growth and cytotoxicity. Flow cytometry was employed to detect cell surface markers of cancer stem cell population. Expression and activity of matrix metalloproteinases were probed with qRT-PCR and zymogen assay. To identify pathways affected by sublethal dose of Dox, exploratory RNAseq was performed and results were verified by qRT-PCR in multiple cell lines (MCF7, ZR75-1 and U-2OS). Regulation of Src Family kinases (SFK) by key players in DNA damage response was assessed by siRNA knockdown along with western blot and qRT-PCR. Dasatinib and siRNA for Fyn and Yes was employed to inhibit SFKs and verify their role in increased migration and invasion in MCF7 cells treated with sublethal doses of Dox. Results The results show that sublethal Dox treatment leads to increased migration and invasion in otherwise non-invasive MCF7 breast cancer cells. Mechanistically, these effects were independent of the epithelial mesenchymal transition, were not due to increased cancer stem cell population, and were not observed with other chemotherapies. Instead, sublethal Dox induces expression of multiple SFK—including Fyn, Yes, and Src—partly in a p53 and ATR-dependent manner. These effects were validated in multiple cell lines. Functionally, inhibiting SFKs with Dasatinib and specific downregulation of Fyn suppressed Dox-induced migration and invasion of MCF7 cells. Conclusions Overall, this study demonstrates that sublethal doses of Dox activate a pro-invasive, pro-migration program in cancer cells. Furthermore, by identifying SFKs as key mediators of these effects, our results define a potential therapeutic strategy to mitigate local invasion through co-treatment with Dasatinib.

Ir(III) Compounds Containing a Terdentate Ligand Are Potent Inhibitors of Proliferation and Effective Antimetastatic Agents in Aggressive Triple-Negative Breast Cancer Cells

“…The treatment of MDA-MB-231 cells with doxorubicin upregulated the expression of N-cadherin (∼3-fold), β-catenin (∼3-fold), and SNAI1 (∼2-fold) and slightly upregulated also the expression of vimentin; in addition, the expression of E-cadherin slightly decreased. These results are consistent and support the view that sub-lethal doses of doxorubicin activate the EMT pathway in breast cancer cells. − Thus, these findings confirm that anticancer chemotherapy using doxorubicin is inappropriate as it can cause drug-resistant TNBC cells to metastasize, so the search for new drugs to treat breast cancer that would show better antitumor effects than doxorubicin is justified.…”

Ozone combined with doxorubicin exerts cytotoxic and anticancer effects on Luminal-A subtype human breast cancer cell line

“…Increased expression of MMP-2 and MMP-9 have been proposed as the prognostic marker in breast cancer 27 . Mohammed et al investigated the effects of sublethal doxorubicin treatment in noninvasive MCF-7 cells and found that doses of 0.6 μM or less of doxorubicin led to increased migration and invasion by increasing the induction and secretion of MMP isoforms, including MMP-2 and MMP-9 28 . The present study showed that 5 μM doxorubicin alone did not change the levels of MMP-2 and MMP-9 in MCF-7 cells, but addition of ozone treatment successfully reduced these levels.…”

The Activity of Novel BCR-ABL Small-Molecule Degraders Containing Pyrimidine Rings and Their Role in Overcoming Drug Resistance

“…e Bcr-abl fusion gene encodes a pathogenic protein (p210BCR-ABL) that continuously increases tyrosine kinase activity and activates multiple downstream signaling pathways to promote the occurrence and development of myeloid leukemia (chronic myeloid leukemia, CML) [25][26][27]. BCR-ABL kinase is closely related to SRCfamily kinases (SFK), which make up the largest nonreceptor tyrosine kinase family in the human body, including LYN, FYN, LCK, HCK, FGR, BLK, YRK, YES, and c-SRC [28][29][30][31][32][33]. ey play a very important role in regulating cell proliferation, differentiation, adhesion, and movement [34][35][36], and abnormal expression of SFK members is related to the occurrence of various tumors in the human body [37][38][39].…”