OBJECTIVE To observe the effects of Yiying Sanjie Formula drug-containing serum on the proliferation and aerobic glycolysis of TPC-1 in human papillary thyroid cancer cells.

METHODS Male SD rats were continuously administered drugs by intragastric gavage for 7 days to prepare the drug-containing serum of Yiying Sanjie Formula. TPC-1 cells in the logarithmic growth phase were taken and divided into 2.5%, 5%, 7.5%, 10% drug-containing serum and blank serum groups. Each group was treated with different concentrations of drug-containing serum and blank serum in 3 replicate wells. The CCK-8 assay was used to detect the cell proliferation at 24, 48, and 72 h after intervention, and the low, medium, and high-dose groups of the drug-containing serum of Yiying Sanjie Formula were selected to carry out subsequent experiments together with the blank serum group. A kit was used to detect the extracellular lactate excretion and the production of adenosine triphosphate(ATP) in cells 48 h after intervention. RT-PCR and Western blotting were used to detect the mRNA and protein expression levels of pyruvate kinase M2(PKM2), hexokinase 2(HK2), lactate dehydrogenase A(LDHA), c-Myc, and hypoxia-inducible factor 1-α(HIF-1α). The visible spectrophotometry was used to detect the expression of the enzyme activities of PK, HK, and LDH.

RESULTS Compared with the blank serum group, the drug-containing serum of Yiying Sanjie Formula could inhibit the proliferation of TPC-1 cells. The 5%, 7.5% and 10% drug-containing serum interventions for 48 h were used as the low, medium and high dose groups for the follow-up experiments. Compared with the blank serum group, cellular lactate efflux and ATP production were significantly reduced in each drug-containing serum group in a dose-dependent manner; mRNA and protein expression of HK2, PKM2, LDHA, HIF-1α and c-Myc were reduced in the high dose group(P<0.05 or P<0.01 or P<0.001); PK, HK and LDH enzyme activities were reduced in the medium and high dose groups(P<0.01 or P<0.001).

CONCLUSION The drug-containing serum of Yiying Sanjie Formula can inhibit the proliferation and aerobic glycolysis of TPC-1 cells, and its mechanism of action may be related to the inhibition of c-Myc/ HIF-1α signaling pathway, down-regulation of PK, HK, LDH expression and enzyme activities, and reduction of lactate and ATP production.