白色念珠菌细胞壁不溶性β-葡聚糖（β-glucan）抗肿瘤细胞的实验研究
发布时间：2018-12-18 07:37
【摘要】： 【目的】白色念珠菌细胞壁不溶性β-葡聚糖(candida albicans insolubleβ-glucan,CAIBG)是一类生物反应调节剂(biological response modifiers,BRM),具有抗肿瘤、升白细胞、抗病毒等广泛的生物学活性。本研究前期实验已经成功地从白色念珠菌细胞壁中分离出不溶性β-葡聚糖,并通过动物实验证实了CAIBG能对抗环磷酰胺(cyclophosphamide,CTX)引起的免疫抑制作用,具有升高小鼠外周血中白细胞数的功能,而且能诱导小鼠外周血中TNF-α、IL-1β的升高以及小鼠脾脏中TNF-αmRNA的表达水平升高,并在基因、分子水平研究了CAIBG的升白细胞作用机理。本实验从白色念珠菌细胞壁提取不溶性β-葡聚糖,观察CAIBG对体外培养的B16恶性黑色素瘤细胞、SGC-7901胃腺癌细胞、SMMC-7721肝癌细胞和XW-05肺腺癌细胞的生长抑制作用并初步探讨其机制。 【方法】首先从白色念珠菌细胞壁提取不溶性β-葡聚糖,体外培养B16恶性黑色素瘤细胞、SGC-7901胃腺癌细胞、SMMC-7721肝癌细胞和XW-05肺腺癌细胞,当细胞处于对数生长期时加入不同浓度的CAIBG,用光镜观察细胞的形态以及生长情况。用改良四甲基偶氮哗盐(简称改良MTT)法测定不同时间和浓度的CAIBG对以上细胞的生长抑制作用;在以上几株肿瘤细胞中选择对CAIBG最敏感的一株,选其在临床上的治疗药物顺铂,比较其与CAIBG对该肿瘤细胞生长抑制作用,再结合流式细胞仪检测加入CAIBG后肿瘤细胞的凋亡率和细胞周期分布时相。 【结果】CAIBG对B16恶性黑色素瘤细胞、SGC-7901胃腺癌细胞和XW-05肺腺癌细胞有不同程度的体外生长抑制作用,生长抑制率最高分别为42.8%、59.88%、61.97%,IC_(50)分别约为1506.5μg/ml、429.8μg/ml、293.4μg/ml;在形态上具有凋亡细胞的形态特征,不同浓度的CAIBG作用于同一肿瘤细胞的抑制作用具有差异性(P＜0.05),不同肿瘤细胞对同一浓度CAIBG的敏感性差异无显著性(P＞0.05),其中XW-05肺腺癌细胞对CAIBG的敏感性最高,CAIBG和顺铂对XWLC-05肺腺癌的体外抑制作用比较,差异有显著性(P＜0.05)。流式细胞仪检测结果显示CAIBG对XW-05肺腺癌细胞的细胞周期没有影响。对SMMC-7721肝癌细胞的体外生长不具有直接抑制作用。 【结论】不同类型肿瘤细胞对同一浓度CAIBG反应性不同,显示治疗个体化的必要性;同一类型肿瘤细胞对不同浓度药物反应性不一,显示选择最佳药物浓度的重要性。CAIBG具有体外直接抑制肿瘤细胞生长的作用,是具有良好的应用开发前景抗肿瘤药物。
[Abstract]:[objective] Candida albicans cell wall insoluble 尾 -glucan (candida albicans insoluble 尾-glucan,CAIBG is a kind of biological response modulator (biological response modifiers,BRM), which has a wide range of biological activities, such as antitumor, leukocyte, antivirus and so on. In this study, insoluble 尾 -glucan was successfully isolated from the cell wall of Candida albicans, and the immunosuppressive effect of CAIBG on cyclophosphamide (cyclophosphamide,CTX) was confirmed by animal experiments. It can increase the number of white blood cells in peripheral blood of mice, and induce the increase of TNF- 伪, IL-1 尾 in peripheral blood and the expression of TNF- 伪 mRNA in spleen of mice. The mechanism of leukocytosis of CAIBG was studied at molecular level. In this study, insoluble 尾 -glucan was extracted from the cell wall of Candida albicans, and the effects of CAIBG on B16 malignant melanoma cells and SGC-7901 gastric adenocarcinoma cells were observed. Growth inhibition of SMMC-7721 hepatoma cells and XW-05 lung adenocarcinoma cells and its mechanism were investigated. [methods] the insoluble 尾 -glucan was extracted from the cell wall of Candida albicans. B16 malignant melanoma cells, SGC-7901 gastric adenocarcinoma cells, SMMC-7721 hepatoma cells and XW-05 lung adenocarcinoma cells were cultured in vitro. When the cells were in logarithmic growth period, different concentrations of CAIBG, were added to observe the morphology and growth of cells by light microscope. The effects of CAIBG at different time and concentration on the growth inhibition of the above cells were determined by modified tetramethylazo salt (MTT) method. One of the tumor cells was selected as the most sensitive to CAIBG and its clinical therapeutic drug cisplatin was selected to compare the inhibitory effect of cisplatin with CAIBG on the growth of the tumor cells. The apoptosis rate and cell cycle distribution phase of tumor cells after adding CAIBG were detected by flow cytometry. [results] CAIBG had different inhibitory effects on B16 malignant melanoma cells, SGC-7901 gastric adenocarcinoma cells and XW-05 lung adenocarcinoma cells in vitro, and the highest growth inhibition rates were 42.8% and 59.88%, respectively. IC_ (50) was about 1506.5 渭 g / ml, 429.8 渭 g / ml and 293.4 渭 g / ml, respectively. Morphological characteristics of apoptotic cells were observed, and the inhibitory effects of different concentrations of CAIBG on the same tumor cells were different (P < 0. 05). There was no significant difference in the sensitivity of different tumor cells to the same concentration of CAIBG (P > 0. 05). The sensitivity of XW-05 lung adenocarcinoma cells to CAIBG was the highest. The inhibitory effects of CAIBG and cisplatin on XWLC-05 lung adenocarcinoma were compared in vitro. The difference was significant (P < 0.05). Flow cytometry showed that CAIBG had no effect on the cell cycle of XW-05 lung adenocarcinoma cells. It has no direct inhibitory effect on the growth of SMMC-7721 hepatoma cells in vitro. [conclusion] different types of tumor cells have different reactivity to the same concentration of CAIBG, which indicates the necessity of individualized treatment. The reactivity of the same type of tumor cells to different concentrations of drugs is different, which indicates the importance of choosing the best concentration of drugs. CAIBG has the function of inhibiting the growth of tumor cells directly in vitro and is a promising antitumor drug.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R379
本文编号：2385570
[Abstract]:[objective] Candida albicans cell wall insoluble 尾 -glucan (candida albicans insoluble 尾-glucan,CAIBG is a kind of biological response modulator (biological response modifiers,BRM), which has a wide range of biological activities, such as antitumor, leukocyte, antivirus and so on. In this study, insoluble 尾 -glucan was successfully isolated from the cell wall of Candida albicans, and the immunosuppressive effect of CAIBG on cyclophosphamide (cyclophosphamide,CTX) was confirmed by animal experiments. It can increase the number of white blood cells in peripheral blood of mice, and induce the increase of TNF- 伪, IL-1 尾 in peripheral blood and the expression of TNF- 伪 mRNA in spleen of mice. The mechanism of leukocytosis of CAIBG was studied at molecular level. In this study, insoluble 尾 -glucan was extracted from the cell wall of Candida albicans, and the effects of CAIBG on B16 malignant melanoma cells and SGC-7901 gastric adenocarcinoma cells were observed. Growth inhibition of SMMC-7721 hepatoma cells and XW-05 lung adenocarcinoma cells and its mechanism were investigated. [methods] the insoluble 尾 -glucan was extracted from the cell wall of Candida albicans. B16 malignant melanoma cells, SGC-7901 gastric adenocarcinoma cells, SMMC-7721 hepatoma cells and XW-05 lung adenocarcinoma cells were cultured in vitro. When the cells were in logarithmic growth period, different concentrations of CAIBG, were added to observe the morphology and growth of cells by light microscope. The effects of CAIBG at different time and concentration on the growth inhibition of the above cells were determined by modified tetramethylazo salt (MTT) method. One of the tumor cells was selected as the most sensitive to CAIBG and its clinical therapeutic drug cisplatin was selected to compare the inhibitory effect of cisplatin with CAIBG on the growth of the tumor cells. The apoptosis rate and cell cycle distribution phase of tumor cells after adding CAIBG were detected by flow cytometry. [results] CAIBG had different inhibitory effects on B16 malignant melanoma cells, SGC-7901 gastric adenocarcinoma cells and XW-05 lung adenocarcinoma cells in vitro, and the highest growth inhibition rates were 42.8% and 59.88%, respectively. IC_ (50) was about 1506.5 渭 g / ml, 429.8 渭 g / ml and 293.4 渭 g / ml, respectively. Morphological characteristics of apoptotic cells were observed, and the inhibitory effects of different concentrations of CAIBG on the same tumor cells were different (P < 0. 05). There was no significant difference in the sensitivity of different tumor cells to the same concentration of CAIBG (P > 0. 05). The sensitivity of XW-05 lung adenocarcinoma cells to CAIBG was the highest. The inhibitory effects of CAIBG and cisplatin on XWLC-05 lung adenocarcinoma were compared in vitro. The difference was significant (P < 0.05). Flow cytometry showed that CAIBG had no effect on the cell cycle of XW-05 lung adenocarcinoma cells. It has no direct inhibitory effect on the growth of SMMC-7721 hepatoma cells in vitro. [conclusion] different types of tumor cells have different reactivity to the same concentration of CAIBG, which indicates the necessity of individualized treatment. The reactivity of the same type of tumor cells to different concentrations of drugs is different, which indicates the importance of choosing the best concentration of drugs. CAIBG has the function of inhibiting the growth of tumor cells directly in vitro and is a promising antitumor drug.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R379
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