Preliminary Study On The Inhibitory Effect Of Baicalin On Ovarian Cancer Cell SKOV3 And Its Mechanis

Ovarian cancer is one of the malignant tumors in female reproductive system with a high incidence and high mortality rate,showing hidden early onset symptom.At present,the main clinical treatment of ovarian cancer is surgical resection,supplemented with radiotherapy and chemotherapy.However,the serious side effects of radiotherapy and chemotherapy are still huge challenges for clinical treatment.Chinese medicine is a traditional treatment in China.It has achieved good therapeutic effects with its unique treatment concept and characteristics of medication.Scutellaria baicalensis is one of the commonly used traditional Chinese medicines.It is mainly used to treat diseases such as jaundice.Baicalin is one of the main active ingredients in Scutellaria baicalensis with a high proportion,which can exert various pharmacological effects such as anti-tumor,anti-inflammation,anti-bacterial and neuroprotective.In this paper,ovarian cancer cell line SKOV3 was used as experimental object to study the inhibitory effect of baicalin on ovarian cancer cell line SKOV3,and the possible pathways and mechanisms of the effects were explored,which may provide some biological basis to study the function of inhibition effects on ovarian cancer cells of baicalin.Experimental method:Experiment 1:The ovarian cancer cell line SKOV3 was divided into the control group and the baicalin group.The cells in control group were treated with DMSO.The cells in baicalin group were treated with 20μM,40μM,80μM,120μM and 160μM baicalin for 48h,and the absorbance of each group was detected by MTS method to observe the effect of baicalin on the proliferation of ovarian cancer cell line SKOV3;Experiment 2:The ovarian cancer cell SKOV3 was divided into the control group and the baicalin group.The cells in control group were added with DMSO.The cells in baicalin group were treated with 80 pM baicalin for 48 hours.The cell cycle distribution was detected by flow cytometry to observe the effect of baicalin on the cell cycle of ovarian cancer cell line SKOV3;Experiment 3:The ovarian cancer cell line SKOV3 was divided into the control group and the baicalin group.The cells in control group were added with DMSO.The cells in baicalin group were treated with 80μM baicalin for 48 hours.The apoptosis rate was detected by flow cytometry to observe the effect of baicalin on the apoptosis in ovarian cancer cell line SKOV3;Experiment 4:The ovarian cancer cell line SKOV3 was divided into the control group and the baicalin group.The cells in control group were added with DMSO.The cells in baicalin group were treated with 80μM baicalin for 48 hours.The expression and phosphorylation of STAT3,p65,AKT,JAK2 and SRC were detected by Western Blot.mRNA and protein expression of Cyclin A and the STAT3 signaling pathway related factor such as Mcl-1,Bcl-xl,Bcl-2,Survivin were detected by real-time fluorescent quantitative PCR and Western Blot.Experimental results:Experiment 1:After treatment with different concentrations of baicalin for 48h,the absorbance of cells in 20μM baicalin group had no significant change,and the absorbance of cells in 40μM-160μM baicalin group decreased significantly(P<0.001).The inhibition rate of the 20μM baicalin group had no significant change,and the inhibition rate of the 40μM-160μM baicalin group increased significantly(P<0.001);Experiment 2:In the control group,the proportion of cells in G1 phase was 57.27%,the proportion of cells in S phase was 34.70%,and the proportion of cells in G2 phase was 8.17%.In baicalin group,the proportion of cells in G1 phase was 39.26%,the proportion of cells in S phase was 60.27%,and the proportion of cells in G2 phase was 0.00%.Compared with the control group,the proportion of cells in G1 phase and G2 phase was significantly decreased after treatment with 80 μM baicalin(P<0.001),while the proportion of cells in S phase was significantly increased(P<0.001);Experiment 3:The proportion of early apoptotic cells in the control group was 9.43%,and the proportion of early apoptotic cells in the baicalin group was 34.47%.Compared with the control group,the early apoptosis of cells increased significantly after 80μM baicalin treatment(P<0.001);Experiment 4:Compared with the control group,after treatment with 80μM baicalin,the mRNA and protein expression of Cyclin A in ovarian cancer cell SKOV3 were increased(P<0.01,P<0.05);STAT3,p65,AKT total protein and AKT phosphorylation had no significant changes;STAT3,p65 phosphorylation expression were decreased(P<0.05);JAK2,SRC total protein and phosphorylation had no significant change;Mcl-1,Bcl-xl,Bcl-2,Survivin mRNA expression were decreased(P<0.001,P<0.001),the expression of Mcl-1,Bcl-xl,Bcl-2,Survivin protein were decreased(P<0.01,P<0.05).Conclusion:Experiment 1:20μM baicalin ovarian cancer cell SKOV3 proliferation has no obvious inhibition effect,40μM-160μM baicalin can significantly inhibit the proliferation of ovarian cancer cell SKOV3;Experiment 2:80μM baicalin can induce S phase arrest in ovarian cancer cell line SKOV3,which may be one of the mechanisms of baicalin inhibition effect on the proliferation of ovarian cancer cells;Experiment 3:80μM baicalin can induce apoptosis of ovarian cancer cell line SKOV3,which may be one of the mechanisms of baicalin inhibition effect on the proliferation of ovarian cancer cells;Experiment 4:The mechanism of baicalin inhibiting the proliferation of ovarian cancer cell SKOV3 and inducing S phase arrest and apoptosis may be through up-regulating Cyclin A expression,inhibiting STAT3 pathway,and then reducing STAT3 downstream genes Mcl-1,Bcl-x1,Bcl-2 and Survivin expression.This experiment provides a new evidence for studying the role and mechanism of baicalin against ovarian cancer.