PMID- 28314283
OWN - NLM
STAT- MEDLINE
DCOM- 20170411
LR  - 20191210
IS  - 1791-7530 (Electronic)
IS  - 0250-7005 (Print)
IS  - 0250-7005 (Linking)
VI  - 37
IP  - 3
DP  - 2017 Mar
TI  - Stable shRNA Silencing of Lactate Dehydrogenase A (LDHA) in Human MDA-MB-231 
      Breast Cancer Cells Fails to Alter Lactic Acid Production, Glycolytic Activity, 
      ATP or Survival.
PG  - 1205-1212
AB  - BACKGROUND: In the US, African Americans have a high death rate from 
      triple-negative breast cancer (TNBC), characterized by lack of hormone receptors 
      (ER, PR, HER2/ERRB2) which are otherwise valuable targets of chemotherapy. There 
      is a need to identify novel targets that negatively impact TNBC tumorigenesis. 
      TNBCs release an abundance of lactic acid, under normoxic, hypoxic and hyperoxic 
      conditions; this referred to as the Warburg effect. Accumulated lactic acid 
      sustains peri-cellular acidity which propels metastatic invasion and malignant 
      aggressive transformation. The source of lactic acid is believed to be via 
      conversion of pyruvate by lactate dehydrogenase (LDH) in the last step of 
      glycolysis, with most studies focusing on the LDHA isoform. MATERIALS AND 
      METHODS: In this study, LDHA was silenced using long-term MISSION(R) shRNA 
      lentivirus in human breast cancer MDA-MB-231 cells. Down-regulation of LDHA 
      transcription and protein expression was confirmed by western blot, 
      immunocytochemistry and qPCR. A number of parameters were measured in fully 
      viable vector controls versus knock-down (KD) clones, including levels of lactic 
      acid produced, glucose consumed, ATP and basic metabolic rates. RESULTS: The data 
      show that lentivirus V-165 generated a knock-down clone most effective in 
      reducing both gene and protein levels to less than 1% of vector controls. Stable 
      KD showed absolutely no changes in cell viability, lactic acid production, ATP, 
      glucose consumption or basic metabolic rate. Given the complete absence of impact 
      on any observed parameter by LDH-A KD and this being somewhat contrary to 
      findings in the literature, further analysis was required to determine why. 
      Whole-transcriptome analytic profile on MDA-MB-231 for LDH subtypes using Agilent 
      Human Genome 4x44k microarrays, where the data show the following component 
      breakdown. Transcripts: 30.47 % LDHA, 69.36% LDHB, 0.12% LDHC and 0.05% LDHD. 
      CONCLUSION: These findings underscore the importance of alternative isoforms of 
      LDH in cancer cells to produce lactic acid, when LDHA is silenced or inhibited. 
      LDHA silencing alone is not effective in hampering or inducing changes in 
      survival, metabolism or lactic acid produced in a cell line with high 
      concentrations of LDHB. Future research will be required to confirm effects of 
      dual LDHA/B knockdown and further confirm that the sole source of lactic acid 
      produced occurs through LDH (all isoforms) in breast cancer cells.
CI  - Copyright(c) 2017, International Institute of Anticancer Research (Dr. George J. 
      Delinasios), All rights reserved.
FAU - Mack, Nzinga
AU  - Mack N
AD  - College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, 
      Tallahassee, FL, U.S.A.
FAU - Mazzio, Elizabeth A
AU  - Mazzio EA
AD  - College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, 
      Tallahassee, FL, U.S.A.
FAU - Bauer, David
AU  - Bauer D
AD  - College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, 
      Tallahassee, FL, U.S.A.
FAU - Flores-Rozas, Hernan
AU  - Flores-Rozas H
AD  - College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, 
      Tallahassee, FL, U.S.A.
FAU - Soliman, Karam F A
AU  - Soliman KF
AD  - College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, 
      Tallahassee, FL, U.S.A. karam.soliman@famu.edu.
LA  - eng
GR  - G12 MD007582/MD/NIMHD NIH HHS/United States
GR  - P20 MD006738/MD/NIMHD NIH HHS/United States
PT  - Journal Article
PL  - Greece
TA  - Anticancer Res
JT  - Anticancer research
JID - 8102988
RN  - 0 (Isoenzymes)
RN  - 0 (RNA, Small Interfering)
RN  - 33X04XA5AT (Lactic Acid)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 1.1.1.27 (L-Lactate Dehydrogenase)
RN  - EC 1.1.1.27.- (Lactate Dehydrogenase 5)
RN  - IY9XDZ35W2 (Glucose)
SB  - IM
MH  - Adenosine Triphosphate/metabolism
MH  - Breast Neoplasms/*enzymology/*genetics
MH  - Cell Line, Tumor
MH  - Cell Survival
MH  - Down-Regulation
MH  - Female
MH  - Gene Expression Regulation, Neoplastic
MH  - *Gene Silencing
MH  - Glucose/metabolism
MH  - Glycolysis
MH  - Humans
MH  - Isoenzymes/genetics/metabolism
MH  - L-Lactate Dehydrogenase/genetics/*metabolism
MH  - Lactate Dehydrogenase 5
MH  - Lactic Acid/*metabolism
MH  - Polymerase Chain Reaction
MH  - RNA, Small Interfering/*genetics
PMC - PMC5363403
MID - NIHMS852544
OTO - NOTNLM
OT  - LDH
OT  - Lactate dehydrogenase A
OT  - glycolysis
OT  - lactic acid
OT  - triple-negative breast cancer
COIS- Conflicts of Interest There are no conflicts of interest.
EDAT- 2017/03/21 06:00
MHDA- 2017/04/12 06:00
PMCR- 2017/03/23
CRDT- 2017/03/19 06:00
PHST- 2017/01/11 00:00 [received]
PHST- 2017/01/17 00:00 [accepted]
PHST- 2017/03/19 06:00 [entrez]
PHST- 2017/03/21 06:00 [pubmed]
PHST- 2017/04/12 06:00 [medline]
PHST- 2017/03/23 00:00 [pmc-release]
AID - 37/3/1205 [pii]
AID - 10.21873/anticanres.11435 [doi]
PST - ppublish
SO  - Anticancer Res. 2017 Mar;37(3):1205-1212. doi: 10.21873/anticanres.11435.