Background Temozolomide (TMZ) is a first-line drug for the treating glioblastoma. polymerase string reaction, and Traditional western blotting. Outcomes Long-term TMZ treatment elevated CIN-mediated genomic diversity in U251TMZ1, U251TMZ2 and T98GTMZ cells but reduced it in C6TMZ and C6R2TMZ cells. U251TMZ1 and U251TMZ2 cell lines, established in parallel with a similar treatment procedure with the only difference in the duration of treatment, underwent individual phenotypic changes. U251TMZ1 experienced a reduced proliferation and Eprosartan mesylate invasion but increased migration, whereas U251TMZ2 experienced an enhanced proliferation and Eprosartan mesylate invasion but no changes in migration. U251TMZ1 and U251TMZ2 cells exhibited individual patterns in expression/activation of transmission transduction proteins (e.g., MDM2, p53, ERK, AKT, and ASK). C6TMZ and C6R2TMZ cells experienced lower proliferation, colony formation efficiency and migration, whereas T98GTMZ cells experienced increased colony formation efficiency without any changes in proliferation, migration, and invasion. TMZ-treated lines exhibited a differential response to a reduction in glucose concentration and an increased resistance to TMZ re-challenge but not temsirolimus (mTOR inhibitor) or U0126 (MEK1/2 inhibitor) treatment. Conclusion Long-term TMZ treatment selected resistant genotype-phenotype variants or generated novel versatile phenotypes by increasing CIN. An increase of resistance to TMZ re-challenge seems to be the only predictable trait intrinsic to all long-term TMZ-treated tumour cells. Changes in genomic diversity may be responsible for heterogeneous phenotypes of TMZ-treated cell lines. Electronic supplementary material The online version of this article (doi:10.1186/s12935-016-0311-8) contains supplementary material, which is available to authorized Eprosartan mesylate users. shows a percentage of metaphases with numbers of chromosomes? 60 or? 90. c The karyotype differences between cell lines were demonstrated by alignment and comparison of karyographs of vehicle- and TMZ-treated derivatives. A list of all CCAs/NCCAs (in the same purchase as depicted in the of karyographs) and their duplicate number variation are available in Extra file 1: Desk S1. d A listing of karyotypic parameters of every cell series. e Chromosomal CTG3a displaying the regions of hereditary gain/loss. in the (in the (of karyographs) and their duplicate number variation are available in Extra file 3: Desk S3. b A listing of karyotypic parameters of every cell series. c Chromosomal displaying the regions of hereditary gain/loss. in the (in the (of karyographs) and their duplicate number variation are available in Extra file 5: Desk S5. e A listing of karyotypic parameters of every cell line. f The chromosome present the certain specific areas of hereditary gain/reduction. Detailed explanation of duplicate number modifications (CNAs) are available in Extra file 6: Desk S6 Temozolomide promotes flexible phenotype adjustments To elucidate how TMZ affected oncogenic features of cells, we analyzed cell proliferation initial. Previous studies confirmed the fact that proliferation of long-term TMZ-treated glioblastoma cells was elevated, reduced or unchanged (Desk?1). U251 cells proliferated quicker than U251TMZ1 cells but slower than U251TMZ2 cells. Simply no difference in proliferation between T98GTMZ and T98G cells Eprosartan mesylate was observed. C6TMZ and C6R2TMZ cells proliferated slower than C6R1 and C6 cells, respectively. Furthermore, C6R1 and C6R2TMZ cells proliferated slower than C6TMZ and C6 cells, respectively (Fig.?4a), recommending the fact that rat mind microenvironment might Eprosartan mesylate choose for slower-dividing C6 cells preferentially. Alternatively, in harvested C6 derivatives vivo, adapted for the various metabolic and growth-stimulating microenvironment within the mind, may undergo stress, when reintroduced to an in vitro culture. Additionally, we cannot exclude an effect of DMSO as it induced cytotoxicity at certain concentrations in vivo [44]. However, much lower DMSO concentration/volume (20?%/200?l) was injected during this study than was previously reported in ([44] and refs therein). Open in a separate windows Fig.?4 Long-term TMZ treatment promotes diverse changes in.
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