ABSTRACT
ObjectiveTo investigate effect of lyophilized powder of modified Huangqi Gancaotang on proliferation， apoptosis， invasion， migration， and epithelial-mesenchymal transition of human non-small cell lung cancer cells （A549， PC9） and possible mechanism. MethodEffect of 1.0， 2.0， 4.0， 8.0， 12.0 g·L-1 modified Huangqi Gancaotang on the proliferation of non-small cell lung cancer cells was detected by cell counting kit-8 （CCK-8） assay. A549 and PC9 cells were classified into the blank group and the low-， medium-， and high-dose Huangqi Gancaotang groups （2.0， 4.0， 8.0 g·L-1）. Plate cloning assay was used to examine the effect of modified Huangqi Gancaotang on cell cloning ability. Hoechst 33342 staining and flow cytometry were employed to detect the apoptosis， and scratch assay and Transwell migration assay were applied to examine cell migration and invasion abilities， respectively. Mammosphere assay was used to examine the sphere-forming ability of tumor cells， and real-time polymerase chain reaction （Real-time PCR） to detect the mRNA expression of stemness-related molecules octamer-binding transcription factor 4 （Oct-4）， human sex-determining region Y-box 2 （Sox2）， and homeobox transcription factor （Nanog） to assess cancer stem cell activity. The protein expression of B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated death promoter （Bad）， Bcl-2-associated X protein （Bax）， cleaved Caspase-3， Caspase-3， E-cadherin， N-cadherin， vimentin， matrix metalloproteinase-2 （MMP-2）， β-catenin， c-Myc， Cyclin D1， and zinc-finger transcription factor （Slug） was determined by Western blot. ResultThe proliferation ability of A549 and PC9 cells was significantly inhibited after 24 h and 48 h treatment with 1.0， 2.0， 4.0， 8.0， and 12.0 g·L-1 lyophilized powder of modified Huangqi Gancaotang compared with that in the blank group and the inhibition was dose- and time-dependent （P<0.05， P<0.01）. Compared with the blank group， the low-， medium-， and high-dose modified Huangqi Gancaotang suppressed the cloning ability of A549 and PC9 cells （P< 0.05， P<0.01）， and high-dose modified Huangqi Gancaotang induced apoptosis of A549 and PC9 cells （P<0.01）. In comparison with the blank group， the low-， medium-， and high-dose modified Huangqi Gancaotang inhibited the invasion and migration of A549 and PC9 cells （P<0.05， P<0.01）. Compared with the blank group， the low-， medium-， and high-dose modified Huangqi Gancaotang significantly decreased volume of the microspheres of A549 cells and the mRNA expression of Oct-4， Sox2， and Nanog in A549 and PC9 cells （P<0.05， P<0.01）. Compared with the blank group， the medium- and high-dose modified Huangqi Gancaotang down-regulated the expression of the anti-apoptotic protein Bcl-2 （P<0.05， P<0.01）， up-regulated the expression of pro-apoptotic proteins Bad， Bax， and cleaved Caspase-3/Caspase-3 （P<0.05， P<0.01） in A549 and PC9 cells， decreased the expression of MMP-2， N-cadherin， and vimentin （P<0.05， P< 0.01）， and raised the E-cadherin expression （P<0.05， P<0.01）. Moreover， the medium-dose and high-dose modified Huangqi Gancaotang all reduced the expression of β-catenin， c-Myc， Cyclin D1， and Slug in A549 and PC9 cells （P<0.01）. ConclusionModified Huangqi Gancaotang can inhibit the proliferation， invasion， migration， activity of cancer stem cells， and epithelial-mesenchymal transition of human non-small cell lung cancer （A549， PC9） cells and induce apoptosis， and the mechanism is the likelihood that it regulates Wnt/β-catenin signaling pathway.
ABSTRACT
BACKGROUND:The aim of present study is to demonstrate that volatile oil of flos magnoliae biondii pamp(VOMbp) possess anti-immunology inflammatory property.OBJECTIVE:To explore the effect of VOMbp on adjuvant arthritis in rats.DESIGN:Grade sampling,randomized controlled quantitative analysis.SETTING:Teaching and Researching Department of Pharmacology,Chengdu University of Traditional Chinese Medicine.MATERIALS:This study was carried out at the Teaching and Researching Department of Pharmacology,Chengdu University of Traditional Chinese Medicine from December to April in 1999. Totally 120 Wistar rats were involved.METHODS :Wistar rats were randomly divided into 4 groups :model control group, 0.28 mL/kg and 0.14 mL/kg VOMbp group,and 25 mg/kg dex-amethasone group. Allergy were induced in all rats through intra-cutaneous injection of Freund's adjuvant (0.1 mL/rat)at metatarsus so as to establish multiple arthritis model in rat. According to different dosage,the experiments were carried out in three batches with 10 rats in each group.Rats in the first batch were given VOMbp three days before inducing allergy for consecutwe 3 days,in order to observe the influence of VOMbp on adjuvant induced acute non-specific inflammatory;Rats in the second batch were given VOMbp on the 8th day sensitization allergy for consecutive 7days so as to explore the protective effect on secondary pathological changes;rats in the third batch were administratedon the 19th day after sensitization for consecutive 7 days for study in therapeutic effect on sec ondary pathological changes. Meanwhile foot tumefaction and the level of prostaglandin E2in ankle arthritis were observed.MAIN OUTCOME MEASURES: ① Effect of VOMbp on the adjuvant induced acute non-specific inflammatory. ② Preventive effect on adjuvant arthritis. ③ Therapeutic effect on adjuvant arthritis.RESULTS :① VOMbp could significantly suppress the primary foot tume faction. ②Moreover VOMbp of preventive dosage could most significantly suppress the secondary arthritis tumefaction [(0.231±0.065)mL of 0.28 mL/kg group and ( 0.206±0.082 ) mL of 0.14 mL/kg group] ,accompanied by PG-E2 decrement in tumefaction tissues [(0.133 2±0.057 2), (0.138 5 ±0.042 6)OD278nm correspondingly].③ No obvious effect of VOMbp of therapeutic dosage on secondary inflammatory tumefatcion of arthritis could be observed in 0.14 mL/kg group,but obvious decrement of prostaglandin E2 could be observed in edema tissue;While 0.28 mL/kg VOMbp exerted most significant or significant influence on above parameters. CONCLUSION:VOMbp possess inhibiting effect on Freund's adjuvant induced primary foot tumefaction and secondary arthritis,with the preventive effect better than therapeutic effects.The therapeutic effect was proved better in higher dosage group.