Experimental Study Of The Magnolol Inhibits Human Pancreatic Cancer Cells Growth And Metastasis

Pancreatic cancer is one of the most malignant and intractable digestive system tumors. More than 80 percent of pancreatic cancer diagnosed on clinical which has already laid in advanced stage and occurred distant metastasis lead to lose the opportunity to surgical treatment. So chemotherapy became one of the most important adjuvant strategies to the pancreatic cancer. However, the effect rate of chemotherapy drugs commonly used is not high, and has some toxic side-effects. Therefore, it is urgent to find a new and effective anti- cancer drug to treat with pancreatic cancer.Magnolol is a hydroxylated biphenyl compound isolated from the stem bark and root of Magnolia officinalis. Honokiol, is Magnolol’s isomer. It is reported that Magnolol have been avarious pharmacological function.Not only had long-lasting central muscle relaxation, CNS inhibition, anti-bacterial, anti-pathogenic microbial, anti-ulcer functions, but also had anti-oxidation, anti-inflammatory, anti-tumor and inhibiting morphine withdrawal response, preventing platelet aggregation and other pharmacological effects. In recent years, studies have shown that magnolia can induce the occurrence of apoptosis in certain human tumor cells. In addition, Autophagy triggered by Magnolol derivative negatively regulates angiogenesis lead to the development of effective therapeutics in apoptosis-resistant cancer. ObjectiveIn order to explore the clinical significance of Magnolol treatment of pancreatic cancer. In this study, we explored the proliferation, apoptosis and cell cycle, and Magnolol antimetastatic activity of four human pancreatic cancer cells after treated with Magnolol in vitro. Futhermore, we also studied the mechanism of Magnolol impacted on anti-pancreatic cancer in vitro.(1) CCK-8 kits was used to detect the proliferation of PANC-1, CFPAC-1, ASPC-1, MIAPa Ca-2 pancreatic cancer cells cell after treated with Magnolol.(2) Hochest test was used to detect the apoptotic bodies of four human pancreatic cancer cells after treated with Magnolol.(3) Flow cytometric was used to detect the apoptosis of four human pancreatic cancer cells after treated with Magnolol.(4) Flow cytometric was used to detect the cells’ cycle of four human pancreatic cancer cells after treated with Magnolol.(5) Cell-base adhesion assay was used to determine the effect of Magnolo on the adhesion of four human pancreatic cancer cells to fibronectin.(6) The scratch test was used to determine the effect of Magnolo on the migration of four human pancreatic cancer cells.(7) Transwell assay was used to determine the effect of Magnolo on the invasion of four human pancreatic cancer cells.(8) Western bloting was used to to detect the protein level of cathepsin D.(9) Caspase-3 with spectrophotometric detection kit was used to to detect cells within Caspase-3 content with different concentrations of Magnolol.(10) Flow cytometric was used to detect the effect of cathepsin D inhibitor(pepstatin A) on the the apoptosis of Magnolol.(11) Caspase-3 with spectrophotometric detection kit used to examine the protein level of Caspase-3 after treated with Magnolol and pepstatin A. Results(1) By CCK-8 method we found Magnolol inhibited four human pancreatic cancer cells’ proliferation and Magnolol inhibited cell viability in a dose- and time-dependent manner.(2) By Hochest33258 staining and flow cytometry we found that Magnolol can induce apoptosis and cell cycle arrest in the S phase of the cells.(3) By cell-base adhesion and scratch test and invasion assay experiments, with the Magnolol Methods concentration increased, we found that cell adhesion ability,Cell migrationand cell invasion capability of four human pancreatic cancer cells were gradually decrease.(4) We used Western bloting to analysis that MIAPa Ca-2 cells treated with different concentrations Magnolol for 24 hours, the expression of cathepsin D levels increased significantly. Meanwhile, MIAPa Ca-2 cells were treated with Magnolol and pepstatin A. Caspase-3 with spectrophotometric detection kit was used to detect the presence of Caspase-3. The expression of Caspase-3 protein in experimental group was same with the control group. These results indicated that Magnolol-Induced Apoptosis in pancreatic cancer Cells was involved in alterations of expression and activity of lysosomal protease Cathepsin D. ConclusionIt was clearly that Magnolol could effectively inhibite the proliferation, adhesion, invasion and metastasis of four human pancreatic cancer cells. Furthermore, the mechanism of Magnolol induced apoptosis via regulating the function of cathepsin D.