益瘿散结方含药血清对甲状腺乳头状癌TPC-1细胞增殖及有氧糖酵解的影响  

Effect of Yiying Sanjie Formula of Drug-containing Serum on Proliferation and Aerobic Glycolysis of Papillary Thyroid Cancer TPC-1 Cells

作　　者：张雪婷 王菊勇[1] ZHANG Xueting;WANG Juyong(Cancer Institute,Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院肿瘤研究所,上海200032

出　　处：《中国现代应用药学》2025年第3期402-409,共8页Chinese Journal of Modern Applied Pharmacy

摘　　要：目的观察益瘿散结方含药血清对人甲状腺乳头状癌细胞TPC-1增殖及有氧糖酵解的影响。方法雄性SD大鼠连续灌胃给药7 d,制备益瘿散结方含药血清。取对数期生长的TPC-1细胞,分为2.5%、5%、7.5%、10%含药血清组和空白血清组,每组3个复孔,分别用不同浓度含药血清和空白血清进行干预。采用CCK-8检测干预24、48、72 h细胞增殖情况,并筛选益瘿散结方含药血清低、中、高剂量组同空白血清组进行后续实验;采用试剂盒检测干预48 h后细胞乳酸外排量和三磷酸腺苷(adenosine triphosphate,ATP)生成量情况;采用RT-PCR和Western blotting检测M2型丙酮酸激酶(pyruvate kinase M2,PKM2)、已糖激酶2(hexokinase 2,HK2)、乳酸脱氢酶A(lactate dehydrogenase A,LDHA)、c-Myc和缺氧诱导因子1α(hypoxia-inducible factor 1-α,HIF-1α)mRNA和蛋白表达情况;采用可见分光光度法检测PK、HK和LDH酶活性表达情况。结果与空白血清组比较,益瘿散结方含药血清可抑制TPC-1细胞增殖,以5%、7.5%和10%含药血清干预48 h作为低、中、高剂量组进行后续实验。与空白血清组比较,各含药血清组细胞乳酸外排量和ATP生成量显著降低,并且呈剂量依赖性;高剂量组HK2、PKM2、LDHA、HIF-1α和c-Myc的mRNA和蛋白表达量降低(P<0.05或P<0.01或P<0.001);中、高剂量组PK、HK和LDH酶活性降低(P<0.01或P<0.001)。结论益瘿散结方含药血清能够抑制TPC-1细胞增殖、抑制细胞有氧糖酵解,其作用机制可能与其抑制c-Myc/HIF-1α信号通路,下调PK、HK、LDH表达及酶活性,减少乳酸和ATP生成有关。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-1αand 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 Form

关 键 词：益瘿散结方 含药血清 甲状腺癌 有氧糖酵解 

分 类 号：R285.5[医药卫生—中药学]