| In recent years,the incidence and mortality of malignant tumors have been increasing in China.On the one hand,the low level of public health census,social prevention and control of cancer propaganda is weak,early screening and diagnosis of low penetration rate,the majority of cancer patients diagnosed in the advanced stage,the treatment effect is poor.On the other hand,the problem of ecological environment is more and more serious.The high incidence of cancer is closely related to air pollution and drinking water safety.Diosmetin is a kind of natural flavonoids.It has many pharmacologic activity such as anti-tumor,anti-inflammatory,anti-oxidation and protection of the central nervous system.Objective: To observe the diosmetin in vitro on five human tumor cell lines and a cell line of rat liver fibrosis growth inhibition,and further explores the possible mechanism of MCF-7.To observe the inhibitory action of diosmetin on hepatoma H22 ascitic tumor,H22 solid tumor and investigate their mechanisms.Methods: The inhibitory effect of different concentration of diosmetin on human breast cancer cells MCF-7,human hepatoma cell HepG2,human gastric cancer cell MGC-803,human prostate cancer cells PC3,human ovarian cancer SK-OV-3 cells were determined by MTTassay.The morphological changes of MCF-7 cells before and after diosmetin effect were observed under inverted microscope.Annexin V-FITC/PI,PI and JC-1 were used as fluoresent probes to measure the apoptosis rate,cell cycle and mitochondrial membrane potential respectively with flow cytometer.The phospho MAPK were detected by Western blot.H22 ascitic cells were inoculated right armpit of mice to construct solid tumor model,the mice that successful tumor types were randomly divided into five groups,which are the model group,positive control group(5-Fu,15mg·kg-1),diosmetin high,medium and low dose group(80,20,5 mg·kg-1).Observe the effect of diosmetin on the weight of hepatoma H22 solid tumor,inhibition rate and the organ index.The H22 ascitic tumor model was established in mice by intraperitoneal injection of 0.2 mL H22 ascitic cells.The animals were subsequently divided into 4 groups randomly,treated with normal saline,5-FU,high dose diosmetin(80 mg·kg-1)and middle dose diosmetin(20 mg·kg-1),respectively.The body weights of the mice were measured every two days and the mouse behavior was monitored every day,survival time was recorded to evaluate life extension effect.Results:(1)MTT results showed that diosmetin had a certain inhibition effect on MCF-7,MGC-803,PC3,HepG2 and SK-OV-3 cell lines.The IC50 values of the four kinds of tumor cells were 12.90±2.67?mol/L,30.94±2.26?mol/L,44.79±2.16?mol/L,31.32±1.84?mol/L,respectively.The inhibitory effect of SK-OV-3 cells was weaker,and the IC50 value was greater than 100 ?mol/L.(2)Observation of MCF-7 cells were treated with diosmetin by inverted microscope,compared with the control group,the proliferation of cells was inhibited,the cell number decreased,and cell shrinkage,cell debris increased.(3)Diosmetin can induce apoptosis of MCF-7 cells,and with diosmetin concentration increased,the apoptosis rate increased.(4)Diosmetin can induce G2/M phase arrest in MCF-7 cells,and block a time related,were arrested in G2/M phase cells proportion of up to 62.1%.(5)Diosmetin can induce mitochondrial membrane potential of MCF-7 cells decreased,and mitochondrial membrane potential decreased in a time related.(6)Compared with the control group,along with the increasing of co-incubating time and concentration of diosmetin,p38 and JNK are activated,the expression of phospho-p38 and phospho-JNK increased,while no significant changes in the expression of phospho-ERK.(7)H22 solid tumor experment: Diosmetin has certain inhibitory effect on H22 solid tumor,low,medium and high dose group inhibition rates on H22 solid tumor were 38.3%,45.9% and 49.6%.Diosmetin also have certain inhibition on immune system of mice,but less side effects than positive drug.(8)H22 ascites tumor experiment: Compared with the model group,diosmetin can prolong the survival period of mice.Life extension rate of middle and high dose group were 35.26% and 16.03%.In addition,diosmetin had a certain ease on mise liver damage caused by H22 ascites tumor.Conclusion: The results showed that diosmetin had considerable anti-tumor avtivity in vitro and vivo.The vitro experiments showed that diosmetin had inhibitory effect on the proliferation of many tumor cells.The mechanisms of cell apoptosis induced by diosmetin may be associated with G2/M block,P38 and JNK signaling pathway.In addition,the animal experiments revealed that diosmetin had good anti-tumor activity in vivo. |
PDF Full Text Request