Reduced levels of methyltransferase DNMT2 sensitize human fibroblasts to oxidative stress and DNA damage that is accompanied by changes in proliferation-related miRNA expression

Abstract

Methyltransferase DNMT2 is suggested to be involved in the regulation of numerous processes, however its biological significance and underlying molecular mechanisms remain elusive. In the present study, we have used WI-38 and BJ human fibroblasts as an in vitro model system to investigate the effects of siRNA-based DNMT2 silencing. DNMT2-depleted cells were found to be sensitive to oxidative stress conditions as judged by increased production of reactive oxygen species and susceptible to DNA damage that resulted in the inhibition of cell proliferation. DNMT2 silencing promoted upregulation of proliferation-related and tumor suppressor miRNAs, namely miR-28-3p, miR-34a-3p, miR-30b-5p, miR-29b-3p, miR-200c-3p, miR-28-5p, miR-379-5p, miR-382-5p, miR-194-5p, miR-193b-3p and miR-409-3p. Moreover, DNMT2 silencing induced cellular senescence and DNMT2 levels were elevated in replicatively senescent cells. Taken together, we found that DNMT2 may take part in the regulation of cell proliferation and longevity in human fibroblasts and speculate that the manipulation of DNMT2 levels that limits cell proliferation may be potentially useful anticancer strategy.

Role of miRNAs induced by oxidized low-density lipoproteins in coronary artery disease: the REGICOR Study

“…individuals [38]. And hsa-miR-193b-3p was associated with production of reactive oxygen species in human fibroblasts [39].…”

“…The copyright holder for this preprint (which was not this version posted August 2, 2019. [37][38][39][40][41][42]. Of those, we were particularly interested in hsa-miR-122-5p, hsa-miR-125a-3p, and hsa-miR-193b-3p, as they were validated by qPCR.…”

Treatment with Modified Extracts of the Microalga Planktochlorella nurekis Attenuates the Development of Stress-Induced Senescence in Human Skin Cells

“…Human foreskin fibroblasts BJ (lot 62341989, catalog number ATCC ® CRL-2522 ™ ) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and human epidermal keratinocytes HEK (lot 3057, catalog number 102-05F and lot 1831898, catalog number C0015C) were obtained from Cell Application Inc. (San Diego, CA, USA) and Thermo Fisher Scientific (Waltham, MA, USA), respectively. Population doubling levels were monitored as described previously [17] and only replicative young cells with high proliferative potential were used. Cells were cultured at 37 • C in a cell culture incubator in the presence of 5% CO 2 .…”

“…A cellular model of hydrogen peroxide-induced senescence was considered, as described previously [17,19]. Briefly, BJ and HEK cells were treated with 100 µM hydrogen peroxide for 2 h, and then water and ethanolic extracts were added for up to 7 days to analyze their protective effects.…”

Plant-Derived Molecules α-Boswellic Acid Acetate, Praeruptorin-A, and Salvianolic Acid-B Have Age-Related Differential Effects in Young and Senescent Human Fibroblasts In Vitro

“…The expression of cyclins, cell cycle regulation-associated genes and inflammation response genes was evaluated using Applied Biosystems StepOnePlus ™ Real-Time PCR System and dedicated Real-Time PCR TaqMan ® Array Plates, namely TaqMan ™ Array, Human Cyclins and Cell Cycle Regulation, Fast 96-well (4418768, Applied Biosystems ™ ) and TaqMan ™ Array, Human Cytokine Network, Fast 96-well (4418769, Applied Biosystems ™ ), respectively, according to the manufacturer's instructions (Thermo Fisher Scientific, Waltham, MA, USA). The expression profiles were created as previously described [45]. All results were compared to control conditions (cells at p14 without treatments).…”