Effect of LDH-A shRNA on the growth, apoptosis and expression of LDH-A in a human pancreatic cancer cell line PANC-1

Abstract

Objective To construct a short hairpin RNA (shRNA) targeting LDH-A, and evaluate the effects on growth, apoptosis and the expression of LDH-A in PANC-1 cells. Method Three shRNAs targeting LDH-A were combined to pGCsilencer vector, and transfected into PANC-1 cells. The expression of LDH-A after transfected by the three shRNAs in pancreatic cancer cell lines PANC-1 was detected by quantitative real time PCR. After tranfected by LDH shRNA-3 with the highest inhibited rate, cell growth was analyzed by MTT assay, apoptosis was detected by flow cytometry. The LDH-A expression was detected by reverse transcription polymerase chain reaction, LDH activity was observed with enzyme cytochemical method. Results The 2-AACt of LDH-A shRNA-3 was (0. 47 ± 0. 02), less than the untransfected pancreatic cancer cell(0. 71 ± 0. 01), the LDH-A shRNA-3 could inhibit the expression of the LDH-A most effectively. The growth of the pancreatic cancer cell was inhibited after 12 h transfected by LDH-A shRNA-3, all the absorbance value of transfected cell in 24 h,36 h,48 h,72 h decreased obviously compared to the untreated pancreatic cancer cell(P <0. 01). The apoptosis rate of the transfected cell increased to 61.74%. The inhibition of LDH-A expression in PANC-1 cells transfected by shRNA-3 was significantly and the activity of LDH reduced. Conclusion LDH-A shRNA inhibits the expression of LDH-A, the proliferation of cancer cells inducing the apoptosis of PANC-1 cells. Key words: Pancreatic neoplasms; Lactate dehydrogenases; Glycolysis; RNA interference