白色念珠菌細胞壁不溶性β-葡聚糖（β-glucan）抗腫瘤細胞的實驗研究
發(fā)布時間：2018-12-18 07:37
【摘要】： 【目的】白色念珠菌細胞壁不溶性β-葡聚糖(candida albicans insolubleβ-glucan,CAIBG)是一類生物反應(yīng)調(diào)節(jié)劑(biological response modifiers,BRM),具有抗腫瘤、升白細胞、抗病毒等廣泛的生物學(xué)活性。本研究前期實驗已經(jīng)成功地從白色念珠菌細胞壁中分離出不溶性β-葡聚糖,并通過動物實驗證實了CAIBG能對抗環(huán)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細胞數(shù)的功能,而且能誘導(dǎo)小鼠外周血中TNF-α、IL-1β的升高以及小鼠脾臟中TNF-αmRNA的表達水平升高,并在基因、分子水平研究了CAIBG的升白細胞作用機理。本實驗從白色念珠菌細胞壁提取不溶性β-葡聚糖,觀察CAIBG對體外培養(yǎng)的B16惡性黑色素瘤細胞、SGC-7901胃腺癌細胞、SMMC-7721肝癌細胞和XW-05肺腺癌細胞的生長抑制作用并初步探討其機制。 【方法】首先從白色念珠菌細胞壁提取不溶性β-葡聚糖,體外培養(yǎng)B16惡性黑色素瘤細胞、SGC-7901胃腺癌細胞、SMMC-7721肝癌細胞和XW-05肺腺癌細胞,當(dāng)細胞處于對數(shù)生長期時加入不同濃度的CAIBG,用光鏡觀察細胞的形態(tài)以及生長情況。用改良四甲基偶氮嘩鹽(簡稱改良MTT)法測定不同時間和濃度的CAIBG對以上細胞的生長抑制作用;在以上幾株腫瘤細胞中選擇對CAIBG最敏感的一株,選其在臨床上的治療藥物順鉑,比較其與CAIBG對該腫瘤細胞生長抑制作用,再結(jié)合流式細胞儀檢測加入CAIBG后腫瘤細胞的凋亡率和細胞周期分布時相。 【結(jié)果】CAIBG對B16惡性黑色素瘤細胞、SGC-7901胃腺癌細胞和XW-05肺腺癌細胞有不同程度的體外生長抑制作用,生長抑制率最高分別為42.8%、59.88%、61.97%,IC_(50)分別約為1506.5μg/ml、429.8μg/ml、293.4μg/ml;在形態(tài)上具有凋亡細胞的形態(tài)特征,不同濃度的CAIBG作用于同一腫瘤細胞的抑制作用具有差異性(P＜0.05),不同腫瘤細胞對同一濃度CAIBG的敏感性差異無顯著性(P＞0.05),其中XW-05肺腺癌細胞對CAIBG的敏感性最高,CAIBG和順鉑對XWLC-05肺腺癌的體外抑制作用比較,差異有顯著性(P＜0.05)。流式細胞儀檢測結(jié)果顯示CAIBG對XW-05肺腺癌細胞的細胞周期沒有影響。對SMMC-7721肝癌細胞的體外生長不具有直接抑制作用。 【結(jié)論】不同類型腫瘤細胞對同一濃度CAIBG反應(yīng)性不同,顯示治療個體化的必要性;同一類型腫瘤細胞對不同濃度藥物反應(yīng)性不一,顯示選擇最佳藥物濃度的重要性。CAIBG具有體外直接抑制腫瘤細胞生長的作用,是具有良好的應(yīng)用開發(fā)前景抗腫瘤藥物。
[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- 偽, IL-1 尾 in peripheral blood and the expression of TNF- 偽 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.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R379
本文編號：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- 偽, IL-1 尾 in peripheral blood and the expression of TNF- 偽 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.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R379
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