British Journal of Cancer

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Altered expression of topoisomerase IIalpha contributes to cross-resistant to etoposide K562/MX2 cell line by aberrant methylation

T Asano, K Nakamura, H Fujii, N Horichi, T Ohmori, K Hasegawa, T Isoe, M Adachi, N Otake and Y Fukunaga

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Figure 1 - Unfortunately we are unable to provide accessible alternative text for this. If you require assistance to access this image, please contact help@nature.com or the author

Figure 1.

(A) Accumulation of MX2 in K562/P, K562/ADM and K562/MX cells. Intracellular accumulation of MX2 in K562/P, K562/ADM and K562/MX2 was measured according to Materials and Methods. Representative data from three independent experiments. Error bar showed s.d. (B) The efflux of MX2 in K562/P, K562/ADM and K562/MX2. The efflux of MX2 in K562/P, K562/ADM and K562/MX2 cells was measured according to Materials and Methods. Representative data from three independent experiments. Error bar showed s.d. (C) The accumulation of ADM in K562/P, K562/ADM and K562/MX cells. Intracellular accumulation of ADM in K562/P, K562/ADM and K562/MX2 was measured according to Materials and Methods. Representative data from three independent experiments. Error bar showed s.d. (D) The efflux of ADM in K562/P, K562/ADM and K562/MX2. The efflux of ADM in K562/P, K562/ADM and K562/MX2 cells was measured according to Materials and Methods. Representative data from three independent experiments. Error bar showed s.d.

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Figure 2.

(A) Expression of topoisomerase IIalpha mRNA in K562/P, K562/MX2 and K562/ADM. Human topoisomerase IIalpha mRNA expression in K562/P, K562/ADM and K562/MX2 was measured. Significantly decreased expression of human topoisomerase IIalpha was observed in K562/MX2 cells. In K562/ADM cells, slightly decreased expression of human topoisomerase IIalpha was observed. Representative data from three independent experiments. (B) Expression of topoisomerase IIalpha protein in K562/P, K562/MX2 and K562/ADM: Decreased expression of human topoisomerase IIalpha protein in K562/MX2 was observed. Representative data from three independent experiments.

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Figure 3.

Topo II-mediated decantenation activity in nuclear extracts from in K562/P, K562/MX2 and K562/ADM cells. Decreased decantenation activity in K562/MX2was observed compared to in K562/P and K562/ADM cells. Representative data from three independent experiments.

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Figure 4.

Methylation status of promoter region in K562/P and K562/MX2 cells by methylation-specific restriction enzyme analysis with or without 5-Aza-2'-deoxycytidine treatment. Aberrant methylation of CpG site was observed in K562/MX2 cells at the proximal promoter region (position: -152) by methylation-specific enzyme analysis (arrow head). PCR products in K562/P cells showed arrow. Representative data from five independent experiments.

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Figure 5.

Topoisomerase IIalpha mRNA expression with 5-Aza-2'-deoxycytidine treatment in K562/P and K562/MX2 cells. Increased expression of topoisomerase IIalpha RNA in K562/MX2 treated with 5-Aza-2'-deoxycytidine was observed. However, increased expression in 5AZ treated K562/MX2 cells was not fully restored compared inK562/P cells. Representative data from six independent experiments.

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Table 1 - IC50 against MX2 and etoposide, adriamycin, and vincristine with or without indomethacin, 5-Aza-2[prime]-deoxycytidine (5AZ) treatment - Unfortunately we are unable to provide accessible alternative text for this. If you require assistance to access this image, please contact help@nature.com or the author

Table 1 - IC50 against MX2 and etoposide, adriamycin, and vincristine with or without indomethacin, 5-Aza-2'-deoxycytidine (5AZ) treatment.

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