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LDH-A silencing suppresses breast cancer tumorigenicity through induction of oxidative stress mediated mitochondrial pathway apoptosis

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Abstract

LDH-A, as the critical enzyme accounting for the transformation from pyruvate into lactate, has been demonstrated to be highly expressed in various cancer cells and its silencing has also been approved relating to increased apoptosis in lymphoma cells. In this study, we intend to investigate the correlation between LDH-A and other clinicopathological factors of breast cancer and whether LDH-A silencing could suppress breast cancer growth, and if so the potential mechanisms. 46 breast cancer specimens were collected to study the relation between LDH-A expression and clinicopathological characteristics including menopause, tumor size, node involvement, differentiation, and pathological subtypes classified by ER, PR, and Her-2. shRNAs were designed and applied to silence LDH-A expression in breast cancer cell lines MCF-7 and MDA-MB-231. The effects of LDH-A reduction on cancer cells were studied by a series of in vitro and in vivo experiments, including cell growth assay, apoptosis evaluation, oxidative stress detection, transmission electron microscopy observation, and tumor formation assay on nude mice. LDH-A expression was found to correlate significantly with tumor size and to be independent for other clinicopathological factors. LDH-A reduction resulted in an inhibited cancer cell proliferation, elevated intracellular oxidative stress, and induction of mitochondrial pathway apoptosis. Meanwhile, the tumorigenic ability of LDH-A deficient cancer cells was significantly limited in both breast cancer xenografts. The Ki67 positive cancer cells were significantly reduced in LDH-A deficiency tumor samples, while the apoptosis ratio was enhanced. Our results suggested that LDH-A inhibition might offer a promising therapeutic strategy for breast cancer.

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Abbreviations

LDH-A:

Lactate dehydrogenase A

ATP:

Adenasine triphosphate

Her-2:

Human epidermal growth factor receptor 2

G6PD:

Glucose-6-phosphate dehydrogenase

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

TKTL1:

Transketolase-like protein 1

NADH:

Nicotinamide adenine dinucleotide reduced disodium salt

TCA:

Tricarboxylic acid

ER:

Estrogen receptor

PR:

Progesterone receptor

shRNA:

Short hairpin RNA

ROS:

Reactive oxygen species

NAC:

N-acetylcysteine

OXPHOS:

Oxidative phosphorylation

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Acknowledgments

We thank Vicki Geall in the Department of Research Service in The University of Hong Kong for editorial assistance. We also appreciate the great help from The Electronic Microscope Unit in The University of Hong Kong. This research was funded by seed fundings from The University of Hong Kong.

Conflict of interest

The authors declare that they have no potential financial competing interests that may in any way gain or lose financially from the publication of this manuscript at present or in the future. Non-financial competing interests are also not involved in the manuscript.

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Authors and Affiliations

  1. School of Chinese Medicine, The University of Hong Kong, Estates Building, 10 Sassoon Road, Hong Kong, China

    Zhi-Yu Wang, Jian-Gang Shen, Neng Wang & Jian-Ping Chen

  2. Unimed Medical Institute, Hong Kong, China

    Tjing Yung Loo

  3. School of Pharmaceutical Science, Sun Yat-sen University, Guangzhou, China

    Dong-Mei Wang & De-Po Yang

  4. Department of Clinical Oncology, The University of Hong Kong, Pokfulam, Hong Kong, China

    Xin-Yuan Guan

  5. The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China

    Sui-Lin Mo

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  1. Zhi-Yu Wang
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Correspondence to Jian-Ping Chen.

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Wang, ZY., Loo, T.Y., Shen, JG. et al. LDH-A silencing suppresses breast cancer tumorigenicity through induction of oxidative stress mediated mitochondrial pathway apoptosis. Breast Cancer Res Treat 131, 791–800 (2012). https://doi.org/10.1007/s10549-011-1466-6

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  • DOI: https://doi.org/10.1007/s10549-011-1466-6

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