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Cisplatin selects short forms of the mitochondrial DNA OriB variant (16184 – 16193 poly-cytosine tract), which confer resistance to cisplatin

View Article: PubMed Central - PubMed

ABSTRACT

A number of alternations in mitochondrial DNA (mtDNA) have been reported in different types of cancers, and the role of mtDNA in cancer has been attracting increasing interest. In order to investigate the relationship between mtDNA alternations and chemosensitivity, we constructed cybrid (trans-mitochondrial hybrid) cell lines carrying a HeLa nucleus and the mtDNA of healthy individuals because of the presence of somatic alternations in the mtDNA of many cancer cells. After a treatment with 1.0 μg/mL cisplatin for 10 days, we isolated 100 cisplatin-resistant clones, 70 of which carried the shorter mtDNA OriB variant (16184–16193 poly-cytosine tract), which was located in the control region of mtDNA. Whole mtDNA sequencing of 10 clones revealed no additional alternations. Re-construction of the HeLa nucleus and mtDNA from cisplatin-resistant cells showed that cisplatin resistance was only acquired by mtDNA alternations in the control region, and not by possible alternation(s) in the nuclear genome.

No MeSH data available.


Related in: MedlinePlus

Scheme for the construction of cybrids and isolation of cisplatin-resistant cells.Since many cancer cell lines have somatic mtDNA alternations, the 9W4 cybrid, which has HeLa nuclear DNA and normal human mtDNA, was used for the cisplatin treatment. In order to evaluate the effects of mtDNA alternations caused by the cisplatin treatment, 9W4 and R13 were enucleated and fused with EB8 neo ρ0 containing a neomycin-resistant gene.
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f1: Scheme for the construction of cybrids and isolation of cisplatin-resistant cells.Since many cancer cell lines have somatic mtDNA alternations, the 9W4 cybrid, which has HeLa nuclear DNA and normal human mtDNA, was used for the cisplatin treatment. In order to evaluate the effects of mtDNA alternations caused by the cisplatin treatment, 9W4 and R13 were enucleated and fused with EB8 neo ρ0 containing a neomycin-resistant gene.

Mentions: Since most cancer cells harbour somatic alternations in mtDNA and these alternations may be associated with resistance to anti-cancer drugs10, we used cybrid cells (A2, 8W5, 9W3, and 9W4) carrying normal mtDNA to isolate cisplatin-resistant cells. These cells (2 × 107 cells each) were exposed to 1.0 μg/mL cisplatin for 10 days and continue to culture in the absence of cisplatin for additional 10 days. The resulting surviving clones were isolated as cisplatin-resistant cells (Fig. 1). We isolated 100 clones (28 clones from 8W5, 27 clones from 9W3, and 45 clones from 9W4), while no clones were obtained from A2 cybrid cells.


Cisplatin selects short forms of the mitochondrial DNA OriB variant (16184 – 16193 poly-cytosine tract), which confer resistance to cisplatin
Scheme for the construction of cybrids and isolation of cisplatin-resistant cells.Since many cancer cell lines have somatic mtDNA alternations, the 9W4 cybrid, which has HeLa nuclear DNA and normal human mtDNA, was used for the cisplatin treatment. In order to evaluate the effects of mtDNA alternations caused by the cisplatin treatment, 9W4 and R13 were enucleated and fused with EB8 neo ρ0 containing a neomycin-resistant gene.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5385546&req=5

f1: Scheme for the construction of cybrids and isolation of cisplatin-resistant cells.Since many cancer cell lines have somatic mtDNA alternations, the 9W4 cybrid, which has HeLa nuclear DNA and normal human mtDNA, was used for the cisplatin treatment. In order to evaluate the effects of mtDNA alternations caused by the cisplatin treatment, 9W4 and R13 were enucleated and fused with EB8 neo ρ0 containing a neomycin-resistant gene.
Mentions: Since most cancer cells harbour somatic alternations in mtDNA and these alternations may be associated with resistance to anti-cancer drugs10, we used cybrid cells (A2, 8W5, 9W3, and 9W4) carrying normal mtDNA to isolate cisplatin-resistant cells. These cells (2 × 107 cells each) were exposed to 1.0 μg/mL cisplatin for 10 days and continue to culture in the absence of cisplatin for additional 10 days. The resulting surviving clones were isolated as cisplatin-resistant cells (Fig. 1). We isolated 100 clones (28 clones from 8W5, 27 clones from 9W3, and 45 clones from 9W4), while no clones were obtained from A2 cybrid cells.

View Article: PubMed Central - PubMed

ABSTRACT

A number of alternations in mitochondrial DNA (mtDNA) have been reported in different types of cancers, and the role of mtDNA in cancer has been attracting increasing interest. In order to investigate the relationship between mtDNA alternations and chemosensitivity, we constructed cybrid (trans-mitochondrial hybrid) cell lines carrying a HeLa nucleus and the mtDNA of healthy individuals because of the presence of somatic alternations in the mtDNA of many cancer cells. After a treatment with 1.0 μg/mL cisplatin for 10 days, we isolated 100 cisplatin-resistant clones, 70 of which carried the shorter mtDNA OriB variant (16184–16193 poly-cytosine tract), which was located in the control region of mtDNA. Whole mtDNA sequencing of 10 clones revealed no additional alternations. Re-construction of the HeLa nucleus and mtDNA from cisplatin-resistant cells showed that cisplatin resistance was only acquired by mtDNA alternations in the control region, and not by possible alternation(s) in the nuclear genome.

No MeSH data available.


Related in: MedlinePlus