| Objective: To detect the express of CDC25A,CDC25B,SMAD3,TGF-β1 in esophageal cancer, approaching the effects of Artesunate against esophageal cancer and related mechanism, searching the new medicine for the therapy of esophagealcancer.Methods1 Researching esophageal cancer,adjacent mucosa to esophageal carcinoma and normal esophageal mucosa at surgical margin of esophageal carcinoma1.1 To study the mRNA of CDC25A,CDC25B,SMAD3,TGF-β1 in 44 example esophageal carcinoma, paired adjacent mucosa to esophageal carcinoma and normal esophageal mucosa at surgical margin of esophageal carcinoma tissues by RT-PCR. The 44 example was all diagnosed by pathology.1.2 To study 58 example primarily esophageal carcinoma and normal esophageal mucosa,to research the cell cycle,apoptosis and the protein of CDC25A,CDC25B,SMAD3.All examples were diagnosed by pathology.2 To study the role of CDC25A,CDC25B,SMAD3,TGF-β1 in the course of Artesunate against esophageal carcinoma. To set up esophageal carcinoma nude mouse animal model, thity in all,divided them for five groups randomly.Every nude mouse injected 6×106.200ul-1 Eca109 cells in left preepipodite, growing tumor in a week ,the mean size of tumor was 0.4mm3, starting to inject drug ,tow weeks ,one course of treatment was one week ,the first group injected Artesunate 100mg.kg-1,the second group injected Artesunate 200mg.kg-1,the third group injected Artesunate 300mg.kg-1,the forth group injected Cisplatin 3mg.kg-1as masculine control group ,the fifth group inject- ted Sodium Chloride as negative control group.injected drugs in abdominal cavity.tow weeks later,killed all of the nude mouse. Taking out of the tumor,divided two share,one share tested cellcycle,Apoptosis and the protein of CDC25A,SMAD3 using Flow cytometry. The other tested the CDC25A,CDC25B,SMAD3 and TGF-β1of the tumor by PT-PCR.GAPDH as internal reference.3 To make use of spss11.5 software carrying out statistical treatment.Using x±s expressed Test data,To compare different groups using ANOVA,To compare two groups by T test.α=0.05 as the size of test.when P<0.05,having statistical significance.Results1 The express of CDC25A,CDC25B,SMAD3 and TGF-β1 mRNA in different pathological changes of esophageal tissues1.1 The express of CDC25A mRNA in different pathological changes of esophageal tissuesThe express of CDC25A mRNA in normal esophageal mucosa,adjacent mucosa to esophageal carcinoma,esophageal carcinoma respectively was 0.426±0.210,0.494±0.205,0.705±0.185,The express of CDC25A mRNA was higher in esophageal carcinoma than others(P<0.05).1.2 The express of CDC25B mRNA in different pathological changes of esophageal tissuesThe express of CDC25B mRNA in normal esophageal mucosa,adjacent mucosa to esophageal carcinoma,esophageal carcinoma respectively was 0.3832±01440,0.4550±0.1744,0.5802±0.4188,The express of CDC25B mRNA was higher in esophageal carcinoma than others(P<0.05).1.3 The express of SMAD3 mRNA in different pathological changes of esophageal tissuesThe express of SMAD3 mRNA in normal esophageal mucosa,adjacent mucosa to esophageal carcinoma,esophageal carcinoma respectively was 0.623±0.281,0.613±0.278,0.479±0.251,The express of SMAD3 mRNA was lower in esophageal carcinoma than others(P<0.05).1.4 The express of TGF-β1 mRNA in different pathological changes of esophageal tissuesThe express of TGF-β1 mRNA in normal esophageal mucosa,adjacent mucosa to esophageal carcinoma,esophageal carcinoma respectively was 0.5963±0.2437,0.5592±0.2575,0.4555±0.1585,The express of TGF-β1 mRNA was lower in esophageal carcinoma than others(P<0.05).2 To study the cell cycle and apoptosis in different pathological changes of esophageal tissues by FCMThe apoptosis of esophageal carcinoma was lower than normal esophageal mucosa(P<0.05). The apoptosis of esophageal carcinoma was (8.10±3.57)%, The apoptosis of normal esophageal mucosa was (9.53±2.26)%. The G1 stage of normal esophageal mucosa was higher than esophageal carcinoma(P<0.05). The G1 stage of normal esophageal mucosa was(86.27±3.72)%,The G1 stage of esophageal carcinoma was(75.07±11.01)%.The proliferation index of normal esophageal mucosa was lower than esophageal carcinoma(P<0.05). The proliferation index of normal esophageal mucosa was(14.49±3.83)%, The proliferation index of esophageal carcinoma was(24.92±11.01)%.3 To study the protein of CDC25A,CDC25B and SMAD3 in different pathological changes of esophageal tissues by FCMThe protein of CDC25A and CDC25B in esophageal carcinoma was higher than normal esophageal mucosa (P<0.05), The protein of SMAD3 in esophageal carcinoma was lower than normal esophageal mucosa(P<0.05).4 The inhibition effects of Artesunate(Art) on the growth of Eca109 transplated subcutaneous tumors in nude miceThe volume and weight of Eca109 transplated subcutaneous tumors in every experiment group was lower than negative control group(P<0.05). The volume of Eca109 transplated subcutaneous tumors in 100mg.kg-1,200mg.kg-1,300mg.kg-1Art group was(0.66±0.29)cm3,(0.28±0.22)cm3,(0.61±0.38)cm3 respectively. The weight of Eca109 transplated subcutaneous tumors in 100mg.kg-1,200mg.kg-1,300mg.kg-1Art group was (0.66±0.27)g,(0.37±0.16)g,(0.65±0.19)g respectively. The volume of Eca109 transplated subcutaneous tumors in DDP group was(0.21±0.07)cm3, The weight of Eca109 transplated subcutaneous tumors in DDP group was (0.33±0.05)g, The volume of Eca109 transplated subcutaneous tumors in normal sodium group was(1.00±0.30)cm3, The weight of Eca109 transplated subcutaneous tumors in normal sodium group was(0.94±0.31)g. The inhibition rate of volume in 100mg.kg-1,200mg.kg-1,300mg.kg-1Art group was 34%,72%,39% respectively, The inhibition rate of weight in 100mg.kg-1,200mg.kg-1,300mg.kg-1Art group was 29.79%,60.64%,30.85% respectively. The inhibition rate of volume in DDP group was 79%,The inhibition rate of weight in DDP group was 64.89%.5 The express of CDC25A,CDC25B,SMAD3,TGF-β1 mRNA in Eca109 transplated subcutaneous tumors in nude mice5.1 The express of CDC25A mRNA100mg.kg-1Art group was 0.8383±0.0454,200mg.kg-1Art group was 0.7300±0.0583,300mg.kg-1Art group was 0.8517±0.0483,DDP group was 0.6850±0.1436,normal sodium group was 0.9650±0.0635. normal sodium group was higher than others(P<0.05). 5.2 The express of CDC25BmRNA100mg.kg-1Art group was 0.5036±0.0041, 200mg.kg-1Art group was 0.4556±0.0353,300mg.kg-1Art group was 0.5024±0.0047,DDP group was 0.4406±0.0354,normal sodium group was 0.5389±0.0407. normal sodium group was higher than others(P<0.05).5.3 The express of SMAD3 mRNA100mg.kg-1Art group was 0.7865±0.0421, 200mg.kg-1Art group was 0.8909±0.0832,300mg.kg-1Art group was 0.7915±0.0582,DDP group was 0.8914±0.0410,normal sodium group was 0.6862±0.0518. normal sodium group was lower than others(P<0.05).5.4 The express of TGF-β1 mRNA100mg.kg-1Art group was 0.6703±0.0193, 200mg.kg-1Art group was 0.7439±0.0546,300mg.kg-1Art group was 0.6777±0.0284,DDP group was 0.7083±0.0628,normal sodium group was 0.6091±0.0405. normal sodium group was lower than others(P<0.05).6 The express of CDC25A,SMAD3 protein in Eca109 transplated subcutaneous tumors in nude miceCDC25A protein: 100mg.kg-1 Art group was 513.5761±18.7518, 200mg.kg-1 Art group was 470.0678±17.2265, 300mg. kg-1Art group was 512.5522±13.3181,DDP group was 484.5414±25.9349,normal sodium group was 552.1977±22.0075. normal sodium group was higher than others(P<0.05). SMAD3 protein: 100mg.kg-1Art group was 418.2193±11.3027, 200mg.kg-1Art group was 454.6632±33.9175, 300mg. kg-1Art group was 418.9937±31.1770,DDP group was 456.5815±37.2966,normal sodium group was 371.4166±23.2261. normal sodium group was lower than others (P<0.05).7 The cell cycle of Eca109 transplated subcutaneous tumors in nude miceThe cell cycle was stopped in G0- G1 stage(66.3%)in Art groups(P<0.05). The S stage in normal sodium group was 42.9%,higher than others,to show that tumor cells grew vigorously.Conclusion1. CDC25A mRNA and protein in esophageal carcinoma was higher than normal esophageal mucosa. CDC25B mRNA and protein in esophageal carcinoma was higher than normal esophageal mucosa.SMAD3 mRNA and protein in esophageal carcinoma was lower than normal esophageal mucosa. TGF-β1 mRNA in esophageal carcinoma was lower than normal esophageal mucosa.these gene and protein maybe the molecule mechanism to cause esophageal carcinoma.2. The apoptosis rate in normal esophageal mucosa was higher than esophageal carcinoma, G1 stage in normal esophageal mucosa was higher than esophageal carcinoma, s stage in esophageal carcinoma was higher than normal esophageal mucosa ,to show that tumor cells grow vigorously.3. Artesunate can stop the growth of Eca109 transplated subcutaneous tumors in nude mice,the middle density Artesunate had the strongest effect in stopping the growth of Eca109 transplated subcutaneous tumors in nude mice in three Artesunate groups.4. Artesunate can stop the transplated subcutaneous tumors cell in G1 stage,so the role of Artesunate against esophageal carcinoma maybe relate to cell cycle blockage.5. Artesunate can increase the expression of SMAD3 and TGF-β1,reduce the expression of CDC25A and CDC25B.so the role of Artesunate against esophageal carcinoma maybe relate to CDC25A,CDC25B,SMAD3 and TGF-β1.
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