Alterations of gene expression and protein synthesis in co-cultured adipose tissue-derived stem cells and squamous cell-carcinoma cells: consequences for clinical applications

Abstract

IntroductionThis is the first study evaluating the interactions of human adipose tissue derived stem cells (ADSCs) and human squamous cell carcinoma cells (SCCs), with regard to a prospective cell-based skin regenerative therapy and a thereby unintended co-localization of ADSCs and SCCs.MethodsADSCs were co-cultured with A431-SCCs and primary SCCs (pSCCs) in a transwell system, and cell-cell interactions were analyzed by assessing doubling time, migration and invasion, angiogenesis, quantitative real time PCR of 229 tumor associated genes, and multiplex protein assays of 20 chemokines and growth factors and eight matrix metalloproteinases (MMPS). Results of co-culture were compared to those of the respective mono-culture.ResultsADSCs’ proliferation on the plate was significantly increased when co-cultured with A431-SCCs (P = 0.038). PSCCs and ADSCs significantly decreased their proliferation in co-culture if cultured on the plate (P <0.001 and P = 0.03). The migration of pSCC was significantly increased in co-culture (P = 0.009), as well as that of ADSCs in A431-SCC-co-culture (P = 0.012). The invasive behavior of pSCCs and A431-SCCs was significantly increased in co-culture by a mean of 33% and 35%, respectively (P = 0.038 and P <0.001). Furthermore, conditioned media from co-cultured ADSC-A431-SCCs and co-cultured ADSCs-pSCCs induced tube formation in an angiogenesis assay in vitro.In A431-SCC-co-culture 36 genes were up- and 6 were down-regulated in ADSCs, in A431-SCCs 14 genes were up- and 8 genes were down-regulated. In pSCCs-co-culture 36 genes were up-regulated in ADSCs, two were down-regulated, one gene was up-regulated in pSCC, and three genes were down-regulated. Protein expression analysis revealed that three proteins were exclusively produced in co-culture (CXCL9, IL-1b, and MMP-7). In A431-SCC-co-culture the concentration of 17 proteins was significantly increased compared to the ADSCs mono-culture (2.8- to 357-fold), and 15 proteins were expressed more highly (2.8- to 1,527-fold) compared to the A431-SCCs mono-culture. In pSCC-co-culture the concentration of 10 proteins was increased compared to ADSCs-mono-culture (2.5- to 77-fold) and that of 15 proteins was increased compared to pSCC mono-culture (2.6- to 480-fold).ConclusionsThis is the first study evaluating the possible interactions of primary human ADSCs with human SCCs, pointing towards a doubtlessly increased oncological risk, which should not be neglected when considering a clinical use of isolated human ADSCs in skin regenerative therapies.

The impact of human adipose tissue-derived stem cells on breast cancer cells: implications for cell-assisted lipotransfers in breast reconstruction

“…High IL-8 levels have been associated with a more advanced tumor stage, an accelerated clinical course with reduced postrelapse survival, and increased lymph node involvement in breast cancer patients [38]. In line with this, it has been shown previously that co-injection of MDA-MB231 and ADSCs leads to significantly higher tumor volumes and increased vessel densities compared to the injection of MDA-MB-231 alone, presumably through higher IL-8 and VEGF levels [10, 12, 20]. We could not detect significant effects of BRCA-ADSC co-culture on cellular proliferation in co-cultures of ADSCs or BRCAs.…”

ADSCs in a fibrin matrix enhance nerve regeneration after epineural suturing in a rat model

“…Another aspect that needs to be addressed in the context of utilization of ADSCs is the potential risk of cancer promotion. Recent pre‐clinical studies highlighted this reasonable concern by demonstrating direct cell‐to‐cell and paracrine effects of ADSCs on skin and breast cancer cells, among others . Proliferation capacity increased leading to a more malignant phenotype associated with higher rates of metastasis and worsened prognosis as well as increased tumor size.…”

“…Recent pre-clinical studies highlighted this reasonable concern by demonstrating direct cell-tocell and paracrine effects of ADSCs on skin and breast cancer cells, among others. [40][41][42] Proliferation capacity increased leading to a more malignant phenotype associated with higher rates of metastasis and worsened prognosis as well as increased tumor size. In contrast, there are several experimental studies showing no or even therapeutic effects of ADSCs on cancer cells of different origin.…”

Adipose Tissue-Derived Stem Cells in Regenerative Medicine

“…The invasive behavior of squamous cell carcinoma cells was significantly increased in co-culture with ADSCs, pointing towards a doubtlessly increased oncological risk [69]. …”