ABSTRACT
ObjectiveTo study the mechanism of Shenbai Jiedu prescription inhibiting the proliferation of HCT116 colorectal cancer （CRC） cells by regulating the phosphatase and tensin homolog deleted on chromosome ten （PTEN）/phosphatidylinositol 3-kinase （PI3K）/ protein kinase B （Akt） signaling pathway. MethodShenbai Jiedu prescription was extracted by water extraction and alcohol precipitation to prepare freeze-dried powder. HCT116 cells were cultured in vitro， and treated with different concentrations of Shenbai Jiedu prescription （2， 4， 8， 16 g·L-1）. The inhibitory effect of Shenbai Jiedu prescription on the proliferation of HCT116 cells was tested by methyl thiazolyl tetrazolium （MTT）. Real-time quantitative PCR was used to detect the mRNA expression levels of PTEN， PI3K， Akt， glycogen synthase kinase-3β （GSK-3β）， c-Myc， survivin and Cyclin D1. Western blot was employed to measure the protein expression levels of PTEN， phosphorylated PTEN （p-PTEN）， PI3K， Akt， phosphorylated Akt （p-Akt）， GSK-3β， phosphorylated GSK-3β （p-GSK-3β）， c-Myc， survivin and Cyclin D1， β-catenin nuclear import was explored by immunofluorescence assay. ResultCompared with the control group， Shenbai Jiedu prescription inhibited the proliferation of HCT116 cells in a dose-dependent manner （P<0.01）. Compared with the control group， the mRNA expression levels of PTEN and GSK-3β were up-regulated whereas those of PI3K， Akt， c-Myc， survivin and CyclinD1 were down-regulated after treatment with Shenbai Jiedu prescription （P<0.01）. The protein expression levels of PTEN， p-PTEN and GSK-3β were up-regulated whereas those of PI3K， Akt， p-Akt， GSK-3β， p-GSK-3β， c-Myc， survivin and CyclinD1 were down-regulated （P<0.05， P<0.01）. Immunofluorescence assay showed that Shenbai Jiedu prescription suppressed β-catenin nuclear import in HCT116 cells. ConclusionShenbai Jiedu prescription inhibited the proliferation of HCT116 cells via the mechanism of regulating the PTEN/PI3K/Akt signaling pathway.
ABSTRACT
Objective To explore the expression and clinical significance of PTEN and CyclinD1 protein in patients with PTC,and to provide theoretic evidence on prognosis evaluation and clinical personalized treatment.Methods 76 patients diagnosed with PCT,treated in Quhua Hospital in Zhejiang Province,from Jan.2015 to Jan.2017,were enrolled.The expression of PTEN and CyclinD1 protein was detected by EnVision two-step immunohistochemical method,and their correlation with clinical pathologic characteristic of PTC was analyzed.Results The negative rate of PTEN of the patients enrolled was 73.68％(56/76),and compared with negative rate of tumor-adjacent tissue(19.74％,15/76),demonstrated statistic significance (x2=44.429,P＜0.05).The down regulation of PTEN expression was positively related with pathological stage,tumor lymph node metastasis and sex (P＜0.05).The positive rate of CyclinD1 was 76.32％(58/76),and compared with positive rate of tumoradjacent tissue(13.16％,10/76),demonstrated statistic significance(x2=61.311,P＜0.05).The up regulation of CyclinD1 protein was positively related with pathological stage and tumor lymph node metastasis (P＜0.05).Conclusions The down regulation of PTEN and up regulation of CyclinD1 are closely related with PTC pathological stage and tumor lymph node metastasis.Detection of PTEN and CyclinD1 would be benefit to early evaluation on prognosis of PTC patients.