| ObjectiveNasopharyngeal carcinoma(nasopharyngeal carcinoma,NPC)is one of the high incidence of malignant tumor in south China,incidence of otolaryngology head of malignant tumor.NPC is a characteristic tumor displaying epidemiological,genetic and regional distribution properties.Although radiotherapy can achieve a high cure rate for early stage NPC,treatment outcomes for the advanced stage are unsatisfactory.Local recurrences and a high frequency of distant metastasis constitute two major causes of treatment failure in NPC after radiotherapy treatment [1].Therefore,novel therapeutic strategies are needed to decrease the incidence and severity associated with this cancer.Pectolinarigenin is an active component of the medicinal plant thistle Eupatorium odoratum.This plant has a wide range of significant applications in clinical medicine and is commonly used as a hemostatic drug.Pectolinarigenin exerts significant cytotoxic activity [2,3].Furthermore,it has shown promise for the treatment of proliferative diseases,such as cancer,and has attracted attention as a candidate compound for cancer chemotherapy.Pectolinarigenin exhibits apoptotic activity,but the mechanism of its apoptotic activity remains unclear.Related articles about inhibitory effect of Pectolinarigenin on nasopharyngeal carcinoma cell had not been reported.Our study used cell experiment and animal experiment to clarify the anti-tumor effect in vivo and in vitro on nasopharyngeal carcinoma cell C666-1,and to further understand its anticancer mechanism,which laid the foundation to deeper study about the anti-tumor effect of Pectolinarigenin on nasopharyngeal carcinoma.Part one Inhibitory effect of Pectolinarigenin on human nasopharyngeal carcinoma cell C666-1 in vitroMethods Compared with CCK 8,clone formation experiment,flow cytometry and Western Blot test,we explored that if pectolinarigenin had a significant inhibitory effect on the proliferation and induction the apoptosis of C666-1 cell,and searched for Nasopharyngeal cancer resistant mechanism of pectolinarigenin.At the same time,ourresearch study on affect C666-1 nasopharyngeal carcinoma(NPC)cell migration of pectolinarigenin by by Transwell migration experiment and scratches migration.Results According to experiment result of CCK 8,C666-1 nasopharyngeal carcinoma(NPC)cell was irradiated with pectolinarigenin which concentration range from2 to 512 micrograms/ml,and set the processing time for 24 hours.Pectolinarigenin treatment group characterized by a dose-response anti-proliferative activity when compared with the control group DMSO.Clone formation assay showed that in a 40 microgramme per milliliter of pectolinarigenin treatment group,number of clones of C666-1 cell was relatively nearly 66.4% lower than in DMSO treatment group(P < 0.01),differences were statistically significant.By flow cytometry instrument testing,compared the result in control group and pectolinarigenin treatment group,we found that pectolinarigenin can accelerate the apoptosis of C666-1 cell.Compared with control group,potential of mitochondria of C666-1 cell which was effect by pectolinarigenin declined obviously,ROS levels intracellular increased after 24 hours.Western blot test results showed that compared with control group,Cleaved Caspase 3,9 protein expression intracellular of pectolinarigenin treatment group increased.In transwell migration assays,the cells that migrated through the pectolinarigenin effects were increased in treatment group when compared with control group DMSO(P<0.05),differences were statistically significant.The scratch test results showed that the migration distance of C666-1 cell in pectolinarigenin treatment group was decreased as the control group(P<0.05),differences were statistically significant.Conclusion In the present study,pectolinarigenin reduced the proliferation,migration and increased apoptosis of C666-1 cells,The inhibitory effect may result from the up-regulated expressions of Cleaved Caspase-3、9 and ROS,The specific mechanisms may relate to the activation of mitochondria signal pathway.Part two Inhibitory effect of Pectolinarigenin on human nasopharyngeal carcinoma C666-1 xenograft in nude miceMethods The model of human nasopharyngeal carcinoma xenograft in nude mice was established and randomly divided into five groups,control group,High-dosePectolinarigenin group,Intermediate-dose Pectolinarigenin group,Low-dose Pectolinarigenin group and DDP group.Drugs were given by intraperitoneal injection daily for 4 weeks.After 4 weeks,nude mice were euthanized and the effect of antitumor on each group was studied.Apoptosis of tumor cells was detected by TUNEL method and H-E staining.The protein expressions of Bax,Bcl-XL and active Caspase-3 in xenografts were examined by Western blotting analysis and immunohistochemistry.The experiment helped to understand the toxic effects of Pectolinarigenin on nude mice.Results Compared with control group(P<0.01),the average tumor volume was much smaller and the average tumor mass was much less in treatment groups.The expressions of Bax and active Caspase-3 were higher(P<0.05)and the expression of Bcl-XL was lower(P<0.05)in treatment groups than in control group.TUNEL result showed the apoptosis index was significantly increased in treatment groups than in control group(P<0.01).The toxicity of Pectolinarigenin to the vital organs(heart,liver and kidney)and blood counts was not obvious.Conclusion Pectolinarigenin can effectively inhibit the growth of human nasopharyngeal carcinoma xenograft in nude mice,and with no obvious adverse effects.The inhibitory effect may result from the induced apoptosis of human nasopharyngeal carcinoma cell and up-regulated expressions of Bax and active Caspase-3,down-regulated expression of Bcl-XL.The specific mechanisms may relate to the activation of mitochondria signal pathway. |
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