ABSTRACT
Objective:To explore the mechanism of promotion effect of juglone combined with cisplatin on the apoptosis of human cervical cancer HeLa cells,and to clarify the effects of its associated signal transduction pathways.Methods:The HeLa cells at logarithmic growth phase were divided into control group,juglone group, cisplatin group and juglone combined with cisplatin group (combined treatment group).The inhibitory rates of proliferation of HeLa cells were detected by MTT assay.The apoptosis was detected by Hoechst 33258 staining. The expressions of Bcl-2, Bax and caspase-3, AKt, and pAKt were detected by Western blotting method. Results:The MTT results showed that the HeLa cell proliferation at 24,48 72 h in each drug group was inhibited;compared with control group,the profileration of HeLa cells in juglone group and cisplatin group was significantly inhibited,especially in combined group. Compared with single drug group,the inhibitory effect in combined treatment group was more significantly.After treatment for 12 h,the typical morphological changes of apoptosis were found in juglone group and cisplatin group by Hoechst 33258 staining,especially in combined treatment group. The Western blotting results showed that the expression levels of Bcl-2 and pAKt in HeLa cells in juglone group and cisplatin group 12 h after treatment were decreased obviously,whereas the expression levels of Bax,Caspase-3,and AKt were increased significantly, especially in combined treatment group compared with control group. Conclusion:Juglone combined with cisplatin could inhibit the PI3K/AKt pathway,thereby promoting the apoptpsis of HeLa cells.
ABSTRACT
AIM:To explore the effect of peptidyl-prolyl cis/trans isomerase (Pin1) inhibitor juglone on apop-tosis of human cervical cancer SiHa cells.METHODS:Cultured SiHa cells were incubated with juglone at concentrations of 10, 20, 50, 80 and 100 μmol/L for 24 h.The SiHa cell activity was detected by methyl thiazolyl tetrazolium ( MTT) assay.The cell apoptosis was analyzed by flow cytometry with Hoechst 33258 staining.The protein levels of cleaved caspase-3,8,9 and PTEN was determined by Western blotting.RESULTS:In different doses of juglone groups, the SiHa cell growth was greatly inhibited ( P<0.05) in a dose-dependent manner as compared with control group.The IC50 of ju-glone was 20.4 μmol/L.After treatment with juglone at concentration of 20 μmol/L for 12 h, the apoptosis of SiHa cells was induced, and the typical morphological changes of cell apoptosis such as karyopyknotic pyknic hyperfluorescence bolus, nuclear fragmentation and apoptotic body were observed by Hoechst 33258 staining.The early apoptotic rate was increased significantly as compared with the control.The protein levels of cleaved caspase-3, 8, 9 and PTEN were also increased sig-nificantly as compared with control group.CONCLUSION:Juglone significantly inhibits the cell activity and induces the apoptosis of SiHa cells in vitro by inhibiting the caspase pathway and increasing the expression of anti-oncogene.
ABSTRACT
BACKGROUND: Considerable studies demonstrate that nitric oxide synthase (NOS)/nitric oxide (NO)plays an important role in maintaining normal function of Sertoli cells, and influences spermatic generation and activation as well as fertilizability.OBJECTIVE: To observe the effect of goat testis extract on NOS activity in Sertoli cells of mice with testis injury caused by heavy mental Pb.DESIGN: Randomized controlled animal trial.SETTING: Department of Histology and Embryology, Jilin Medical College.MATERIALS: This trial was carried out in the laboratory of Histology and Embryology, Jilin Medical College (Key laboratory of the general logistics department of P.L.A) during March 2004 to August 2005. Thirty healthy Kunming male mice were involved and randomized into 3 groups,with 10 in each group: control group, testis injury model group (model group) and goat testis extract-treated group (treatment group).METHODS: The mice in the model group and experimental group were daily administrated with 100 g/L lead acetate, 0.2 mL/(mouse·d), 5 times/wk within 2 weeks, then withdrawal for 1 week. Simultaneously, the mice in the treatment group were subcutaneously injected with goat testis extract at 0.5 mL/(mouse ·d). The mice in the control group were given redistilled water of the same dose as that in the treatment group. After being poisoned fully, the mice were fasted for 12 hours and weighted, finally sacrificed by decapitation. Bilateral testis were dissected, immediately weighted, fixed with formalin and sliced. The NOS changes in Sertoli cells of mice in each group were observed with reduced nicotinamide-adenine dinucleotide phophate-diaphorase(NADPH-d) histochemical method combined with microscope image.MAIN OUTCOME MEASURES: Body mass, bilateral testis mass and NOS absorbance (A) in Sertoli cells of mice in each group after contamination expiration.RESULTS: All the 30 mice were involved in the result analysis, without deletion. ①After contamination expiration, the body mass, bilateral testis mass and NOS A value in treatment group were significantly than those in the model group [(22.47±3.49) g vs. (19.13±3.46) g;(0.113±0.021 ) g vs.(0.089±0.017) g; 0.236±0.020 vs. 0.146±0.023, t=2.151-3.314,P ＜ 0.05-0.01]; ② In the model group, NOS positive Sertoli cells swelled and degenerated; The morphology of NOS positive Sertoli cells in the treatment group was close to that in the control group.CONCLUSION: Goat testis extract can boost the NOS activity in Sertoli cells of mice with testis injury caused by heavy mental pliumbum and has some repairing and protective effect on testis injury, which can provide new thinking for treatment of male sterility.