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Overexpression of lactate dehydrogenase-A in human intrahepatic cholangiocarcinoma: its implication for treatment.

Yu Y, Liao M, Liu R, Chen J, Feng H, Fu Z - World J Surg Oncol (2014)

Bottom Line: Previous studies have shown that lactate dehydrogenase-A (LDH-A) is strongly expressed in several malignancies, that LDH-A expression is associated with poor prognosis, and that LDH-A inhibition severely diminishes tumorigenicity.Reduction of LDH-A by RNA interference (RNAi) inhibited cell growth and induced apoptosis in HuCCT-1 cells.This result correlated with the elevation of cytoplasmic reactive oxygen species (ROS) levels.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Minimally Invasive Surgery, Department of General Surgery, First Affiliated Hospital of Nanjing Medical University, NO, 300, Guangzhou Road, Nanjing 210029, China. fuzan2011@163.com.

ABSTRACT

Background: Previous studies have shown that lactate dehydrogenase-A (LDH-A) is strongly expressed in several malignancies, that LDH-A expression is associated with poor prognosis, and that LDH-A inhibition severely diminishes tumorigenicity. However, little is known about the implications of LDH-A expression in intrahepatic cholangiocarcinoma. The purpose of this study was to investigate the expression of LDH-A and to clarify its effect on intrahepatic cholangiocarcinoma.

Methods: We studied the expression of LDH-A in tissue samples from patients with intrahepatic cholangiocarcinoma (n = 54) using the ultrasensitive surfactant protein (S-P) immunohistochemical method. We then inhibited LDH-A using small hairpin RNA (shRNA) in the cholangiocarcinoma cell line HuCCT-1 in vitro to study the role it plays in promoting growth and escaping apoptosis.

Results: We report that LDH-A was overexpressed in 52 of 54 (96%) paraffin-embedded cancer tissue samples and 0 of 54 para-carcinoma tissue samples. Reduction of LDH-A by RNA interference (RNAi) inhibited cell growth and induced apoptosis in HuCCT-1 cells. This result correlated with the elevation of cytoplasmic reactive oxygen species (ROS) levels.

Conclusions: LDH-A expression is closely correlated with histopathological variables of intrahepatic cholangiocarcinoma, indicating that LDH-A may serve as a new treatment target.

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Related in: MedlinePlus

Cell death was determined by flow cytometry of annexin V- and 7-AAD-stained cells at 72 hours post-transfection with shLDH-A (RNAi) or siControl (NC). The histogram (right) represents the average percentage (±SD) of dead cells. The number of dead cells treated with shLDH-A (RNAi) compared with the control group (NC) has a P value of <0.05 using Student’s t test. 7-ADD, 7-aminoactinomycin D; RNAi, RNA interference; SD, standard deviation; sh, small hairpin.
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Figure 4: Cell death was determined by flow cytometry of annexin V- and 7-AAD-stained cells at 72 hours post-transfection with shLDH-A (RNAi) or siControl (NC). The histogram (right) represents the average percentage (±SD) of dead cells. The number of dead cells treated with shLDH-A (RNAi) compared with the control group (NC) has a P value of <0.05 using Student’s t test. 7-ADD, 7-aminoactinomycin D; RNAi, RNA interference; SD, standard deviation; sh, small hairpin.

Mentions: To investigate the effect of aberrant LDH-A expression in cholangiocarcinoma, RNAi experiments were performed using the HuCCT-1 cell line. We examined the expression of LDH-A at its mRNA and protein levels by performing RT-PCR and western blotting analyses in the HuCCT-1 cell line (Figure 2A, B). The CCK-8 assay showed that the proliferation rates of the HuCCT-1 LDH-A knocked-down cell line (RNAi group) were slower than those of the negative control HuCCT-1 cell lines (NC group) (Figure 3). The absorbance value of transfected cells at 72 and 96 hours decreased markedly compared to the negative control cells (P <0.01). The apoptosis rates of the RNAi group (25.89% ± 1.86%) obviously increased compared to those of the NC group (7.47% ± 2.02%) (Figure 4). Collectively, inhibition of LDH-A resulted in a decrease in the rate of proliferation of HuCCT-1 cells, while apoptosis rates increased. These results implicate the role of LDH-A in tumorigenesis.


Overexpression of lactate dehydrogenase-A in human intrahepatic cholangiocarcinoma: its implication for treatment.

Yu Y, Liao M, Liu R, Chen J, Feng H, Fu Z - World J Surg Oncol (2014)

Cell death was determined by flow cytometry of annexin V- and 7-AAD-stained cells at 72 hours post-transfection with shLDH-A (RNAi) or siControl (NC). The histogram (right) represents the average percentage (±SD) of dead cells. The number of dead cells treated with shLDH-A (RNAi) compared with the control group (NC) has a P value of <0.05 using Student’s t test. 7-ADD, 7-aminoactinomycin D; RNAi, RNA interference; SD, standard deviation; sh, small hairpin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4230420&req=5

Figure 4: Cell death was determined by flow cytometry of annexin V- and 7-AAD-stained cells at 72 hours post-transfection with shLDH-A (RNAi) or siControl (NC). The histogram (right) represents the average percentage (±SD) of dead cells. The number of dead cells treated with shLDH-A (RNAi) compared with the control group (NC) has a P value of <0.05 using Student’s t test. 7-ADD, 7-aminoactinomycin D; RNAi, RNA interference; SD, standard deviation; sh, small hairpin.
Mentions: To investigate the effect of aberrant LDH-A expression in cholangiocarcinoma, RNAi experiments were performed using the HuCCT-1 cell line. We examined the expression of LDH-A at its mRNA and protein levels by performing RT-PCR and western blotting analyses in the HuCCT-1 cell line (Figure 2A, B). The CCK-8 assay showed that the proliferation rates of the HuCCT-1 LDH-A knocked-down cell line (RNAi group) were slower than those of the negative control HuCCT-1 cell lines (NC group) (Figure 3). The absorbance value of transfected cells at 72 and 96 hours decreased markedly compared to the negative control cells (P <0.01). The apoptosis rates of the RNAi group (25.89% ± 1.86%) obviously increased compared to those of the NC group (7.47% ± 2.02%) (Figure 4). Collectively, inhibition of LDH-A resulted in a decrease in the rate of proliferation of HuCCT-1 cells, while apoptosis rates increased. These results implicate the role of LDH-A in tumorigenesis.

Bottom Line: Previous studies have shown that lactate dehydrogenase-A (LDH-A) is strongly expressed in several malignancies, that LDH-A expression is associated with poor prognosis, and that LDH-A inhibition severely diminishes tumorigenicity.Reduction of LDH-A by RNA interference (RNAi) inhibited cell growth and induced apoptosis in HuCCT-1 cells.This result correlated with the elevation of cytoplasmic reactive oxygen species (ROS) levels.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Minimally Invasive Surgery, Department of General Surgery, First Affiliated Hospital of Nanjing Medical University, NO, 300, Guangzhou Road, Nanjing 210029, China. fuzan2011@163.com.

ABSTRACT

Background: Previous studies have shown that lactate dehydrogenase-A (LDH-A) is strongly expressed in several malignancies, that LDH-A expression is associated with poor prognosis, and that LDH-A inhibition severely diminishes tumorigenicity. However, little is known about the implications of LDH-A expression in intrahepatic cholangiocarcinoma. The purpose of this study was to investigate the expression of LDH-A and to clarify its effect on intrahepatic cholangiocarcinoma.

Methods: We studied the expression of LDH-A in tissue samples from patients with intrahepatic cholangiocarcinoma (n = 54) using the ultrasensitive surfactant protein (S-P) immunohistochemical method. We then inhibited LDH-A using small hairpin RNA (shRNA) in the cholangiocarcinoma cell line HuCCT-1 in vitro to study the role it plays in promoting growth and escaping apoptosis.

Results: We report that LDH-A was overexpressed in 52 of 54 (96%) paraffin-embedded cancer tissue samples and 0 of 54 para-carcinoma tissue samples. Reduction of LDH-A by RNA interference (RNAi) inhibited cell growth and induced apoptosis in HuCCT-1 cells. This result correlated with the elevation of cytoplasmic reactive oxygen species (ROS) levels.

Conclusions: LDH-A expression is closely correlated with histopathological variables of intrahepatic cholangiocarcinoma, indicating that LDH-A may serve as a new treatment target.

Show MeSH
Related in: MedlinePlus