消痰散结方干预胃癌裸鼠模型的蛋白质组学研究
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摘要
研究目的：以人胃癌裸鼠皮下移植瘤模型为研究对象,观察消痰散结方对荷瘤裸鼠生存质量及肿瘤生长的影响；运用蛋白质组学技术,观察和分析消痰散结方干预后荷瘤裸鼠血清及肿瘤组织蛋白质组表达的变化,筛选出与消痰散结方作用相关的差异表达蛋白质并对其进行质谱鉴定,探讨消痰散结方对胃癌的作用机制。
研究方法：
1、复制模型：将52只裸鼠随机分为造模组和正常组,其中造模组42只,正常组10只。造模组通过皮下移植人胃癌MKN-45瘤块建立胃癌裸鼠模型。观察成瘤率,待大部分移植瘤体积接近80mm3左右时,剔除移植瘤生长过快或过慢的裸鼠12只,剩余30只荷瘤裸鼠和正常组10只裸鼠进入下一步实验。
2、观察消痰散结方对荷瘤裸鼠生存质量及肿瘤生长的影响：将进入实验的荷瘤裸鼠随机分为模型组、阳性对照组和干预组3组,每组10只。模型组给予生理盐水,阳性对照组给予卡培他滨片(希罗达,Xeloda)混悬液,干预组给予消痰散结方。给药途径均为灌胃,剂量均为0.4ml/只/d,给药时间为2周。密切观察荷瘤裸鼠给药后的反应,并于第0、5、10、15天间隔测量每只裸鼠的瘤径大小及体重。2周后处死全部裸鼠,取血清及肿瘤组织保存备用。
3、消痰散结方对荷瘤裸鼠血清蛋白质组表达的影响：对实验第一部分获得的血清样品进行处理(除去高丰度蛋白)后进行蛋白定量、等电聚焦(IEF)、胶条平衡、SDS-PAGE、染色脱色等步骤获得血清蛋白2-DE凝胶图谱。应用PDQUST图像分析软件将正常组、模型组和干预组3组血清2-DE凝胶图谱进行匹配,将不同胶图匹配点相对含量差异2倍及其以上的点选为差异表达蛋白质点。
4、消痰散结方对荷瘤裸鼠肿瘤组织蛋白质组表达的影响：对实验第一部分获得的肿瘤组织样品进行处理后进行蛋白定量、IEF、胶条平衡、SDS-PAGE、染色脱色等步骤获得瘤组织蛋白2-DE凝胶图谱,应用PDQUST图像分析软件对模型组和干预组2组瘤组织的2-DE胶图进行匹配,不同胶图匹配点相对含量差异2.5倍及其以上的点选为差异表达蛋白质点。
5、差异表达蛋白质的质谱鉴定：在第二、三部分实验中获得的差异表达蛋白质斑点进行胶内酶解,利用MALDI-TOF/TOF-MS质谱仪对差异表达蛋白进行鉴定,并对获得的肽质量指纹图谱(PMF)在相关数据库内进行搜索,确定差异表达蛋白质的身份。
结果：
1、复制模型：实验成功复制了裸鼠人胃癌MKN-45皮下移植瘤模型,成瘤率高达到100%。荷瘤裸鼠通过筛选达到了很好的均一性,保证了后续实验的进行。
2、消痰散结方对荷瘤裸鼠生长的影响：荷瘤裸鼠分组给药后,与模型组和干预组相比,阳性对照组裸鼠出现进食及活动减少、皮肤无光泽、消瘦等症状。给药第5天,模型组、阳性对照组和干预组3组间体重、瘤体积、相对瘤体积差异均无统计学意义(P>0.05)。给药第10天,阳性对照组裸鼠体重较模型组和干预组下降明显(P<0.05)；阳性对照组和干预组裸鼠瘤体积及相对瘤体积较模型组均下降明显(P<0.05)。实验结束后处死所有裸鼠,阳性对照组与干预组的肿瘤生长抑制率分别为48.4%和41.9%。
3、消痰散结方对荷瘤裸鼠血清蛋白质组表达的影响：实验中去除血清中高丰度蛋白质后获得重复性、稳定性较好的血清蛋白质2-DE图谱,该图显示的蛋白斑点在分布、背景等方面基本一致。通过分析软件比对,与正常组比较,在模型组中找到25个差异表达蛋白点,其中12个表达上调,13个表达下调；与模型组比较,在干预组中找到19个差异表达蛋白点,其中14个表达上调,5个表达下调；3组间比较(干预组与模型组、模型组与正常组间比较差异有统计学意义,而干预组与正常组间比较差异无统计学意义),共找到差异表达蛋白点9个,其中与正常组比较,在模型组中表达上调,在干预组表达回调至正常组水平的3个；与正常组比较,在模型组中表达下调,在干预组表达回复至正常组水平的有6个。
4、消痰散结方对荷瘤鼠肿瘤组织蛋白质组表达的影响：肿瘤组织双向电泳凝胶重复性、稳定性较好,显示的蛋白斑点在分布、背景等方面基本一致。模型组与干预组比对,找到41个差异表达蛋白质斑点。其中新生蛋白质斑点11个,8个仅在模型组中出现,3个仅在干预组中出现。在其余30个差异表达蛋白质斑点中,13个表达上调,17个表达下调。
5、差异蛋白质的质谱鉴定：本实验成功鉴定出21个差异表达蛋白质(载脂蛋白A1、超氧化物歧化酶、过氧化物酶1、热休克蛋白27、泛素蛋白连接酶、结合珠蛋白、热休克蛋白60等)。这些蛋白质按其功能特性可分为细胞增殖、分化与凋亡、细胞周期相关蛋白、能量代谢相关酶类、脂质代谢相关蛋白、细胞骨架和运动相关蛋白、氧化应激相关蛋白、热休克相关蛋白等8类蛋白质。
结论：
1、希罗达和消痰散结方对荷瘤裸鼠肿瘤的生长均具有明显的抑制作用,但希罗达对裸鼠存在一定的毒性,消痰散结方能够改善裸鼠的生存状态。
2、消痰散结方对载脂蛋白A1、过氧化物酶1、超氧化物歧化酶、热休克蛋白27等表达有上调作用；对结合珠蛋白、泛素蛋白连接酶、热休克蛋白60、组蛋白甲基转移酶等表达有下调作用。
3、消痰散结方的抗胃癌作用可能与调节细胞增殖、分化与凋亡、细胞周期、能量代谢酶类、脂质代谢、细胞骨架和运动、氧化应激和热休克等相关蛋白质的表达有关。
4、消痰散结方抗胃癌作用具有多靶点、多途径、多环节、多层次的特点。
5、21种差异表达蛋白质可作为肿瘤痰证的物质基础,为肿瘤痰证理论提供了实验数据。
Objective:To observe the impact on tumor growth of tumor-bearing nude mice intervened by XiaotanSanjie Recipe, based on the human gastric cancer mice model. Observe and analyze the serum and tumor tissue protein spectrum changes of tumor-bearing mice intervened by XiaotanSanjie Recipe on the platform of proteomics. Screen the different protein spots related with XiaotanSanjie Recipe and identify some of them to investigate possible anti-cancer mechanism of XiaotanSanjie Recipe.
Methods:
1.Modelling:Fifty-two BALB/c-nu/nu mice were randomly divided into experimental group (n=42), and control group (n=10). MKN-45, a cell line from poor differentiated human gastric cancer, was cultured in vitro to an exponential phase. Then, the cancer cells were injected into the subcutaneous site of BALB/c-nu/nu mice to build tumor-bearing models. When diameters of subcutaneous transplanted tumors grow near 1cm, the mice were sacrificed and active tumor was divided into many smaller pieces with a diameter of 1.5mm. The smaller pieces were inoculated into new BALB/c-nu/nu mice for passage culture in vivo. The cancer cells were activated by the passage culture in vivo 5 times. Then, they were regarded as the original tumor and inoculated again into 42 BALB/c-nu/nu mice to prepare the tumor-bearing models in a large scale with the same method mentioned above. When the volumes of most of transplanted tumors had growed near 80mm3,12 mice models with irregular tumors were excluded.Thirty tumor-bearing mice were remained in the following study.
2. Impact of XiaotanSanjie Recipe on tumor-bearing mice and tumor growth:Thirty tumor-bearing mice were randomly divided into model group, positive control group and intervention group, ten for each. All mice were fed freely. Additionally the the model group was fed with saline, the positive control group received Xeloda and the intervention group was administrated with XiaotanSanjie Recipe. All drugs including saline were delivered by intragastric administration with a dosage of 0.4ml every mouse per day. The response of tumor-bearing mice to treatment was observed carefully, the diameter of tumor and body weight of every mouse was measured every 5 days to draw curves of tumor volume and body weight. All mice were sacrificed after 2 weeks treatment. Their blood was collected from the orbital venous plexus. Draw the supernate blood serum for cryopreservation after centrifugalization. The tumor of every nude mouse model was separated from the mouse and weighted. Some of them were fixed with formalin solution for histopathological examination and others were saved by liquid nitrogen freezing.
3. Impact of XiaotanSanjie Recipe on blood serum proteome expression of tumor-bearing mice:Collect the serum samples obtained from the Part 1 experiment to process protein quantify, isoelectric focusing (IEF), strips balance, SDS-PAGE, stain bleaching after depletion the high abundance proteins in order to get two-dimensional gel electrophoresis maps. Gel maps were analyzed with PDQUST. Points from control group, model group and intervention group were matched and twice or more different content were considered different points.
4. Impact of XiaotanSanjie Recipe on tumor Proteome expression of tumor-bearing mice:Collect the serum samples obtained from the Part 1 experiment to process isoelectric focusing (IEF), strips balance, SDS-PAGE, stain bleaching in order to get two-dimensional gel electrophoresis maps. Gel maps were analyzed with PDQUST. Points from model group and intervention group were matched and 2.5 times or more different content were considered different points.
5. Identification and database search of different proteins:Those different proteins obtained from the Part2 Part3 experiment were digested, analyzed with MALDI-TOF/TOF-MS mass spectrograph and searched from protein database.
Results:
1.Modelling:Tumor-bearing models were successfully established by injecting human gastric cancer MKN-45 injected into the subcutaneously into BALB/c-nu/nu mice. The rate of tumor formation reached 100%. The mice models achieved good homogeneity by excluded the ones bearing irregular tumor, this ensured the following experiments.
2. Impact of XiaotanSanjie Recipe on tumor-bearing nude mice:After administration, mice positive control group appears to lose weight, eat and active less, their skin became dull, compared with model group and intervention group. After administrated for 5 days, there was no significant difference of body weight tumor volume and relative tumor volume in model group, positive control group and intervention group (P＞0.05).After administrated for 10 days, body weight of positive control group decreased significantly compared with model group and intervention group (P<0.05), tumor volume and relative tumor volume of positive control group and intervention group decreased significantly compared with model group (P＜0.05). After the experiment, all the nude mice were sacrificed. Inhibition rate of positive control group and intervention group were 48% and 41% respectively.
3. Impact of XiaotanSanjie Recipe on serum Proteome expression of tumor-bearing mice:Depletion the high abundance proteins the serum can get protein map with satisfying repeatability and stability. The protein points had approximately same distribution and background. By comparison to control group,25 different protein spots were obtained in model group. In those different proteins, the expressions of 12 proteins were promoted and 9 proteins depressed. By comparison to model group,19 different protein spots were obtained in intervention group. In these different proteins, the expression of 14 proteins was promoted and 5 depressed. There were 9 proteins which had significant difference between intervention group and model group, but had no difference between intervention group and control group. Three of them were promoted in model group comparing with control group, but depressed to the normal level in the intervention group. The other six proteins were depressed in model group comparing with control group, but promoted to the normal level in the intervention group.
4. Impact of XiaotanSanjie Recipe on tumor proteome expression of tumor-bearing mice:Two-dimensional gel electrophoresis on samples of tumor tissue had satisfying repeatability and stability. The proteins had a proximately the same distribution and background. By comparison, the model group with the intervention group, a total of 41 different protein spots obtained. In which,8 proteins only appeared in model group,3 only appeared in intervention group. The expression of 30 different proteins reached 2.5 times or more. The expressions of 13 proteins were promoted, and 17 depressed in the intervension group.
5. Identification different proteins and retrieval recognition:A total of 21 most differentially expressed proteins were identified, including APO1, SOD, POX1, HSP27, ubiquity protein ligase, haptoglobin, and HSP60, etc. These proteins can be divided according to their functions into 8 species such as cell proliferation, differentiation or apoptosis related,energy metabolism related, lipid metabolism related,cytoskeleton and sports related, oxidative stress related and heat shock related proteins.
Conclusion:
1. Both Xelod and XiaotanSanjie Recipe inhibited the tumor growth of tumor-bearing nude mice obviously. But Xeloda had certain toxicity in nude mice; XiaotanSanjie Recipe can improve the living condition of nude mice.
2. XiaotanSanjie Recipe can promote the expression level of APO1, POX1, SOD and HSP27.It also can depress the expression level of haptoglobin, ubiquity protein ligase, histone methyltransferase, etc.
3. The study found that XiaotanSanjie Recipe maybe play an anti-tumor effects through regulating the expression of cell proliferation,differentiation or apoptosis related,energy metabolism related, lipid metabolism related,cytoskeleton and sports related, oxidative stress-related and heat shock related proteins.
4. The anti-tumor function of XiaotanSanjie Recipe has the characteristic of multi-target, multi-way, multi-link and multi-level.
5. Proteome study may open up new avenues for Efficacy monitoring and mechanism discovery of traditional Chinese herbal compound.
研究方法：
1、复制模型：将52只裸鼠随机分为造模组和正常组,其中造模组42只,正常组10只。造模组通过皮下移植人胃癌MKN-45瘤块建立胃癌裸鼠模型。观察成瘤率,待大部分移植瘤体积接近80mm3左右时,剔除移植瘤生长过快或过慢的裸鼠12只,剩余30只荷瘤裸鼠和正常组10只裸鼠进入下一步实验。
2、观察消痰散结方对荷瘤裸鼠生存质量及肿瘤生长的影响：将进入实验的荷瘤裸鼠随机分为模型组、阳性对照组和干预组3组,每组10只。模型组给予生理盐水,阳性对照组给予卡培他滨片(希罗达,Xeloda)混悬液,干预组给予消痰散结方。给药途径均为灌胃,剂量均为0.4ml/只/d,给药时间为2周。密切观察荷瘤裸鼠给药后的反应,并于第0、5、10、15天间隔测量每只裸鼠的瘤径大小及体重。2周后处死全部裸鼠,取血清及肿瘤组织保存备用。
3、消痰散结方对荷瘤裸鼠血清蛋白质组表达的影响：对实验第一部分获得的血清样品进行处理(除去高丰度蛋白)后进行蛋白定量、等电聚焦(IEF)、胶条平衡、SDS-PAGE、染色脱色等步骤获得血清蛋白2-DE凝胶图谱。应用PDQUST图像分析软件将正常组、模型组和干预组3组血清2-DE凝胶图谱进行匹配,将不同胶图匹配点相对含量差异2倍及其以上的点选为差异表达蛋白质点。
4、消痰散结方对荷瘤裸鼠肿瘤组织蛋白质组表达的影响：对实验第一部分获得的肿瘤组织样品进行处理后进行蛋白定量、IEF、胶条平衡、SDS-PAGE、染色脱色等步骤获得瘤组织蛋白2-DE凝胶图谱,应用PDQUST图像分析软件对模型组和干预组2组瘤组织的2-DE胶图进行匹配,不同胶图匹配点相对含量差异2.5倍及其以上的点选为差异表达蛋白质点。
5、差异表达蛋白质的质谱鉴定：在第二、三部分实验中获得的差异表达蛋白质斑点进行胶内酶解,利用MALDI-TOF/TOF-MS质谱仪对差异表达蛋白进行鉴定,并对获得的肽质量指纹图谱(PMF)在相关数据库内进行搜索,确定差异表达蛋白质的身份。
结果：
1、复制模型：实验成功复制了裸鼠人胃癌MKN-45皮下移植瘤模型,成瘤率高达到100%。荷瘤裸鼠通过筛选达到了很好的均一性,保证了后续实验的进行。
2、消痰散结方对荷瘤裸鼠生长的影响：荷瘤裸鼠分组给药后,与模型组和干预组相比,阳性对照组裸鼠出现进食及活动减少、皮肤无光泽、消瘦等症状。给药第5天,模型组、阳性对照组和干预组3组间体重、瘤体积、相对瘤体积差异均无统计学意义(P>0.05)。给药第10天,阳性对照组裸鼠体重较模型组和干预组下降明显(P<0.05)；阳性对照组和干预组裸鼠瘤体积及相对瘤体积较模型组均下降明显(P<0.05)。实验结束后处死所有裸鼠,阳性对照组与干预组的肿瘤生长抑制率分别为48.4%和41.9%。
3、消痰散结方对荷瘤裸鼠血清蛋白质组表达的影响：实验中去除血清中高丰度蛋白质后获得重复性、稳定性较好的血清蛋白质2-DE图谱,该图显示的蛋白斑点在分布、背景等方面基本一致。通过分析软件比对,与正常组比较,在模型组中找到25个差异表达蛋白点,其中12个表达上调,13个表达下调；与模型组比较,在干预组中找到19个差异表达蛋白点,其中14个表达上调,5个表达下调；3组间比较(干预组与模型组、模型组与正常组间比较差异有统计学意义,而干预组与正常组间比较差异无统计学意义),共找到差异表达蛋白点9个,其中与正常组比较,在模型组中表达上调,在干预组表达回调至正常组水平的3个；与正常组比较,在模型组中表达下调,在干预组表达回复至正常组水平的有6个。
4、消痰散结方对荷瘤鼠肿瘤组织蛋白质组表达的影响：肿瘤组织双向电泳凝胶重复性、稳定性较好,显示的蛋白斑点在分布、背景等方面基本一致。模型组与干预组比对,找到41个差异表达蛋白质斑点。其中新生蛋白质斑点11个,8个仅在模型组中出现,3个仅在干预组中出现。在其余30个差异表达蛋白质斑点中,13个表达上调,17个表达下调。
5、差异蛋白质的质谱鉴定：本实验成功鉴定出21个差异表达蛋白质(载脂蛋白A1、超氧化物歧化酶、过氧化物酶1、热休克蛋白27、泛素蛋白连接酶、结合珠蛋白、热休克蛋白60等)。这些蛋白质按其功能特性可分为细胞增殖、分化与凋亡、细胞周期相关蛋白、能量代谢相关酶类、脂质代谢相关蛋白、细胞骨架和运动相关蛋白、氧化应激相关蛋白、热休克相关蛋白等8类蛋白质。
结论：
1、希罗达和消痰散结方对荷瘤裸鼠肿瘤的生长均具有明显的抑制作用,但希罗达对裸鼠存在一定的毒性,消痰散结方能够改善裸鼠的生存状态。
2、消痰散结方对载脂蛋白A1、过氧化物酶1、超氧化物歧化酶、热休克蛋白27等表达有上调作用；对结合珠蛋白、泛素蛋白连接酶、热休克蛋白60、组蛋白甲基转移酶等表达有下调作用。
3、消痰散结方的抗胃癌作用可能与调节细胞增殖、分化与凋亡、细胞周期、能量代谢酶类、脂质代谢、细胞骨架和运动、氧化应激和热休克等相关蛋白质的表达有关。
4、消痰散结方抗胃癌作用具有多靶点、多途径、多环节、多层次的特点。
5、21种差异表达蛋白质可作为肿瘤痰证的物质基础,为肿瘤痰证理论提供了实验数据。
Objective:To observe the impact on tumor growth of tumor-bearing nude mice intervened by XiaotanSanjie Recipe, based on the human gastric cancer mice model. Observe and analyze the serum and tumor tissue protein spectrum changes of tumor-bearing mice intervened by XiaotanSanjie Recipe on the platform of proteomics. Screen the different protein spots related with XiaotanSanjie Recipe and identify some of them to investigate possible anti-cancer mechanism of XiaotanSanjie Recipe.
Methods:
1.Modelling:Fifty-two BALB/c-nu/nu mice were randomly divided into experimental group (n=42), and control group (n=10). MKN-45, a cell line from poor differentiated human gastric cancer, was cultured in vitro to an exponential phase. Then, the cancer cells were injected into the subcutaneous site of BALB/c-nu/nu mice to build tumor-bearing models. When diameters of subcutaneous transplanted tumors grow near 1cm, the mice were sacrificed and active tumor was divided into many smaller pieces with a diameter of 1.5mm. The smaller pieces were inoculated into new BALB/c-nu/nu mice for passage culture in vivo. The cancer cells were activated by the passage culture in vivo 5 times. Then, they were regarded as the original tumor and inoculated again into 42 BALB/c-nu/nu mice to prepare the tumor-bearing models in a large scale with the same method mentioned above. When the volumes of most of transplanted tumors had growed near 80mm3,12 mice models with irregular tumors were excluded.Thirty tumor-bearing mice were remained in the following study.
2. Impact of XiaotanSanjie Recipe on tumor-bearing mice and tumor growth:Thirty tumor-bearing mice were randomly divided into model group, positive control group and intervention group, ten for each. All mice were fed freely. Additionally the the model group was fed with saline, the positive control group received Xeloda and the intervention group was administrated with XiaotanSanjie Recipe. All drugs including saline were delivered by intragastric administration with a dosage of 0.4ml every mouse per day. The response of tumor-bearing mice to treatment was observed carefully, the diameter of tumor and body weight of every mouse was measured every 5 days to draw curves of tumor volume and body weight. All mice were sacrificed after 2 weeks treatment. Their blood was collected from the orbital venous plexus. Draw the supernate blood serum for cryopreservation after centrifugalization. The tumor of every nude mouse model was separated from the mouse and weighted. Some of them were fixed with formalin solution for histopathological examination and others were saved by liquid nitrogen freezing.
3. Impact of XiaotanSanjie Recipe on blood serum proteome expression of tumor-bearing mice:Collect the serum samples obtained from the Part 1 experiment to process protein quantify, isoelectric focusing (IEF), strips balance, SDS-PAGE, stain bleaching after depletion the high abundance proteins in order to get two-dimensional gel electrophoresis maps. Gel maps were analyzed with PDQUST. Points from control group, model group and intervention group were matched and twice or more different content were considered different points.
4. Impact of XiaotanSanjie Recipe on tumor Proteome expression of tumor-bearing mice:Collect the serum samples obtained from the Part 1 experiment to process isoelectric focusing (IEF), strips balance, SDS-PAGE, stain bleaching in order to get two-dimensional gel electrophoresis maps. Gel maps were analyzed with PDQUST. Points from model group and intervention group were matched and 2.5 times or more different content were considered different points.
5. Identification and database search of different proteins:Those different proteins obtained from the Part2 Part3 experiment were digested, analyzed with MALDI-TOF/TOF-MS mass spectrograph and searched from protein database.
Results:
1.Modelling:Tumor-bearing models were successfully established by injecting human gastric cancer MKN-45 injected into the subcutaneously into BALB/c-nu/nu mice. The rate of tumor formation reached 100%. The mice models achieved good homogeneity by excluded the ones bearing irregular tumor, this ensured the following experiments.
2. Impact of XiaotanSanjie Recipe on tumor-bearing nude mice:After administration, mice positive control group appears to lose weight, eat and active less, their skin became dull, compared with model group and intervention group. After administrated for 5 days, there was no significant difference of body weight tumor volume and relative tumor volume in model group, positive control group and intervention group (P＞0.05).After administrated for 10 days, body weight of positive control group decreased significantly compared with model group and intervention group (P<0.05), tumor volume and relative tumor volume of positive control group and intervention group decreased significantly compared with model group (P＜0.05). After the experiment, all the nude mice were sacrificed. Inhibition rate of positive control group and intervention group were 48% and 41% respectively.
3. Impact of XiaotanSanjie Recipe on serum Proteome expression of tumor-bearing mice:Depletion the high abundance proteins the serum can get protein map with satisfying repeatability and stability. The protein points had approximately same distribution and background. By comparison to control group,25 different protein spots were obtained in model group. In those different proteins, the expressions of 12 proteins were promoted and 9 proteins depressed. By comparison to model group,19 different protein spots were obtained in intervention group. In these different proteins, the expression of 14 proteins was promoted and 5 depressed. There were 9 proteins which had significant difference between intervention group and model group, but had no difference between intervention group and control group. Three of them were promoted in model group comparing with control group, but depressed to the normal level in the intervention group. The other six proteins were depressed in model group comparing with control group, but promoted to the normal level in the intervention group.
4. Impact of XiaotanSanjie Recipe on tumor proteome expression of tumor-bearing mice:Two-dimensional gel electrophoresis on samples of tumor tissue had satisfying repeatability and stability. The proteins had a proximately the same distribution and background. By comparison, the model group with the intervention group, a total of 41 different protein spots obtained. In which,8 proteins only appeared in model group,3 only appeared in intervention group. The expression of 30 different proteins reached 2.5 times or more. The expressions of 13 proteins were promoted, and 17 depressed in the intervension group.
5. Identification different proteins and retrieval recognition:A total of 21 most differentially expressed proteins were identified, including APO1, SOD, POX1, HSP27, ubiquity protein ligase, haptoglobin, and HSP60, etc. These proteins can be divided according to their functions into 8 species such as cell proliferation, differentiation or apoptosis related,energy metabolism related, lipid metabolism related,cytoskeleton and sports related, oxidative stress related and heat shock related proteins.
Conclusion:
1. Both Xelod and XiaotanSanjie Recipe inhibited the tumor growth of tumor-bearing nude mice obviously. But Xeloda had certain toxicity in nude mice; XiaotanSanjie Recipe can improve the living condition of nude mice.
2. XiaotanSanjie Recipe can promote the expression level of APO1, POX1, SOD and HSP27.It also can depress the expression level of haptoglobin, ubiquity protein ligase, histone methyltransferase, etc.
3. The study found that XiaotanSanjie Recipe maybe play an anti-tumor effects through regulating the expression of cell proliferation,differentiation or apoptosis related,energy metabolism related, lipid metabolism related,cytoskeleton and sports related, oxidative stress-related and heat shock related proteins.
4. The anti-tumor function of XiaotanSanjie Recipe has the characteristic of multi-target, multi-way, multi-link and multi-level.
5. Proteome study may open up new avenues for Efficacy monitoring and mechanism discovery of traditional Chinese herbal compound.
引文
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