Atg13 gene overexpression induces chemotherapy resistance in triple-negative breast cancer cells

1LIU Yong-bo; 2; 3DENG Li; 1GUO Yi-hui; 1LI Yan-ping

doi:10.16689/j.cnki.cn11-9349/r.2019.03.008

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Electronic Journal of Metabolism and Nutrition of Cancer ›› 2019, Vol. 6 ›› Issue (3) : 310-315. DOI: 10.16689/j.cnki.cn11-9349/r.2019.03.008

Atg13 gene overexpression induces chemotherapy resistance in triple-negative breast cancer cells

1LIU Yong-bo，2，3DENG Li， 1GUO Yi-hui，1LI Yan-ping

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Abstract

Objective To explore the impact of gene 13(Atg13) on promoting chemotherapy resistance in triple-negative breast cancer (TNBC) cells. Methods We developed a doxorubicin-resistant MDA-MB-231/Dox cell lines using intermittent application of doxorubicin (Dox). MDA-MB-231/Dox was determined by the drug-sensitive assay and the apoptosis assay (TUNEL staining).The expression of LC3A and LC3B in TNBC cells (MDA-MB-231 and MDA-MB-231/Dox) was detected by Western Blot (WB). The number of LC3B fluorescence spots in the cells was quantified by immunofluorescence. Recombinant plasmid sh-Atg13 was transfected into MDA-MB-231/Dox by liposome， then stable cells (referred to as MDA-MB-231/Dox+sh-Atg13) were obtained by puromycin screening (PM). The Atg13 expression level in MDA-MB-231/Dox+sh-Atg13 and empty plasmid MDA-MB-231/Dox+control plasmid by WB. Changes in autophagy activity were determined by the level of LC3A， LC3B expression and the number of LC3B fluorescent spots. Drug sensitivity of each cell line to Dox was also examined. Results Dox-resistant TNBC cell line MDA-MB-231/Dox were successfully established in vitro. The results showed that the level of basal autophagy in drug-resistant cells was significantly higher than that in sensitive cells MDA-MB-231(P<0.05). After inhibiting the expression of Atg13 gene by recombinant interference plasmid sh-Atg13， the protein expression of autophagy marker LC3B in drug-resistant cells MDA-MB-231/Dox was significantly decreased (P<0.05)， the drug sensitivity of MDA-MB-231/Dox cells to Dox was increased (IC50 value was significant， P<0.05). Conclusion In TNBC cell MDA-MB-231， Atg13 increased chemotherapy resistance by inducing autophagy were determined.

Key words

Triple-negative breast cancer / Autophagy / Atg13 / Chemotherapy resistance

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1LIU Yong-bo，2，3DENG Li， 1GUO Yi-hui，1LI Yan-ping. Atg13 gene overexpression induces chemotherapy resistance in triple-negative breast cancer cells[J]. Electronic Journal of Metabolism and Nutrition of Cancer. 2019, 6(3): 310-315 https://doi.org/10.16689/j.cnki.cn11-9349/r.2019.03.008

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