石蒜堿抑制肺癌NCI-H460細胞增殖及Lewis肺癌小鼠腫瘤生長作用研究
發(fā)布時間：2019-04-13 20:53
【摘要】：目的:通過研究石蒜堿在體外對人肺癌NCI-H460細胞增殖的抑制作用及對NCI-H460細胞凋亡的影響,和體內(nèi)對Lewis肺癌小鼠的抑瘤作用,探討石蒜堿抗腫瘤機制,旨在為臨床非小細胞肺癌的治療尋找新的高效、低毒的化療藥物。方法:1.石蒜堿對NCI-H460的細胞毒作用(1)MTT法檢測石蒜堿抑制NCI-H460細胞增殖的作用,(2)用流式細胞儀檢測NCI-H460細胞的凋亡率,(3)比色法檢測凋亡相關(guān)因子Caspase 3的活性;2.石蒜堿對Lewis肺癌小鼠的抑瘤作用(1)Lewis肺癌細胞株的培養(yǎng),(2)小鼠Lewis肺癌移植瘤模型的建立,(3)處死小鼠,計算抑瘤率,脾臟指數(shù)及胸腺指數(shù),(4)HE染色后觀察細胞形態(tài)。結(jié)果:1.石蒜堿作用NCI-H460細胞24 h后,顯微鏡下觀察細胞形態(tài),經(jīng)石蒜堿作用后,NCI-H460細胞外形不規(guī)則,細胞皺縮,大小不一,細胞破碎程度隨給藥劑量增加而增大,細胞數(shù)量逐漸減少。2.用石蒜堿處理NCI-H460細胞24 h后,,NCI-H460細胞增殖受到明顯的抑制,與對照組相比差異顯著(p0.05,p0.01),并呈現(xiàn)一定的濃度相關(guān)性。IC50為5.79±0.11μmol/L,表明石蒜堿能夠有效抑制肺癌NCI-H460細胞增殖。3.隨著石蒜堿處理濃度的升高,處理組細胞凋亡率逐漸升高,與生理鹽水組相比差異顯著(p0.01)。最高濃度16μmol/L時,凋亡率達到46.2±3.46%。上述結(jié)果表明石蒜堿對NCI-H460細胞的凋亡具有誘導(dǎo)效應(yīng),并呈現(xiàn)一定的劑量依賴性。4.隨著石蒜堿濃度的增加,Caspase 3的活性逐漸增高。石蒜堿處理組與生理鹽水對照組比較,Caspase 3的活性顯著升高(p0.01)。結(jié)果表明石蒜堿可增強肺癌NCI-H460細胞的Caspase 3的活性,從而誘導(dǎo)NCI-H460細胞凋亡。5.與生理鹽水組比較,石蒜堿低、中低、中、中高、高劑量組對Lewis肺癌小鼠腫瘤生長均有抑制作用,其中以高劑量組(40 mg/kg)的作用最為顯著,抑瘤率56.8%(p0.01)。6.荷瘤小鼠實體瘤組織的HE染色實驗結(jié)果顯示生理鹽水組腫瘤細胞生長均勻緊密,核固縮較少,無壞死或點狀壞死;石蒜堿各劑量組及環(huán)磷酰胺組腫瘤細胞可見大量核固縮,核碎裂,腫瘤細胞數(shù)量減少,排列疏松,尤其以高劑量組(40 mg/kg)的作用最為顯著,腫瘤細胞明顯散在分布,數(shù)量明顯減少。結(jié)論:石蒜堿在體外能有效地抑制NCI-H460細胞增殖并促進其凋亡,其機制可能與激活了Caspase 3的表達從而誘導(dǎo)腫瘤細胞凋亡有關(guān);石蒜堿在體內(nèi)亦能有效抑制小鼠Lewis肺癌細胞生長。
[Abstract]:Objective: to study the inhibitory effect of Lycorine on the proliferation of human lung cancer NCI-H460 cells in vitro and the apoptosis of NCI-H460 cells, and the antitumor effect of Lycorine on Lewis lung cancer mice in vivo, and to explore the anti-tumor mechanism of garlicine. The aim of this study is to find new effective and low toxic chemotherapeutic drugs for the treatment of clinical non-small cell lung cancer (NSCLC). Methods: 1. Cytotoxicity of allicin to NCI-H460 (1) the inhibitory effect of allicin on the proliferation of NCI-H460 cells was detected by MTT assay; (2) the apoptosis rate of NCI-H460 cells was detected by flow cytometry; (3) the activity of apoptosis-related factor Caspase-3 was detected by colorimetry; 2. Antitumor effect of allicin on Lewis lung cancer mice (1) culture of Lewis lung cancer cell line, (2) establishment of transplanted tumor model of Lewis lung cancer in mice, (3) killing mice, calculation of tumor inhibition rate, spleen index and thymus index, (4) the morphology of the cells was observed after HE staining. Results: 1. After treated with allicin for 24 h, the morphology of NCI-H460 cells was observed under microscope. After treated with garlicine, the shape of NCI-H460 cells was irregular, the cells shrank, the size of the cells was different, and the degree of cell fragmentation increased with the increase of dosages. The number of cells gradually decreased. 2. When NCI-H460 cells were treated with garlicine for 24 h, the proliferation of NCI-H460 cells was significantly inhibited (p0.05, p0.01), and the IC50 was 5.79 鹵0.11 渭 mol / L, compared with the control group (p0.05, p0.01), and the IC50 was 5.79 鹵0.11 渭 mol / L, and the IC50 was 5.79 鹵0.11 渭 mol / L. It is suggested that allicin can effectively inhibit the proliferation of lung cancer NCI-H460 cells. 3. With the increase of the concentration of allicin, the apoptosis rate of the treated group increased gradually, and the difference was significant compared with that of the saline group (p0.01). At the highest concentration of 16 渭 mol / L, the apoptosis rate reached 46.2 鹵3.46%. These results suggest that Lycorine can induce apoptosis of NCI-H460 cells in a dose-dependent manner. 4. With the increase of the concentration of allicin, the activity of Caspase-3 increased gradually. Compared with the control group, the activity of Caspase-3 was significantly increased in the garlicine treatment group (p0.01). The results showed that allicin could enhance the activity of Caspase-3 in lung cancer NCI-H460 cells and induce the apoptosis of NCI-H460 cells. 5. Compared with normal saline group, Lycorine had inhibitory effect on tumor growth of Lewis lung cancer mice in low, middle and high dose groups, among which the high dose group (40 mg/kg) had the most significant effect. The tumor inhibition rate was 56.8% (p0.01). The results of HE staining in solid tumor tissue of tumor-bearing mice showed that the tumor cells in the saline group grew uniformly and closely with less nuclear condensation and no necrosis or punctate necrosis. A large number of nuclear pyknosis, nuclear fragmentation, decrease in the number of tumor cells and loose arrangement were observed in various dose groups and cyclophosphamide groups, especially in the high dose group (40 mg/kg), and the distribution of tumor cells was obvious. There was a marked decrease in the number. Conclusion: garlicine can effectively inhibit the proliferation and promote the apoptosis of NCI-H460 cells in vitro. The mechanism may be related to the activation of Caspase-3 expression and the induction of apoptosis of tumor cells. Allicin can also inhibit the growth of Lewis lung cancer cells in vivo.
【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R734.2
本文編號：2457938
[Abstract]:Objective: to study the inhibitory effect of Lycorine on the proliferation of human lung cancer NCI-H460 cells in vitro and the apoptosis of NCI-H460 cells, and the antitumor effect of Lycorine on Lewis lung cancer mice in vivo, and to explore the anti-tumor mechanism of garlicine. The aim of this study is to find new effective and low toxic chemotherapeutic drugs for the treatment of clinical non-small cell lung cancer (NSCLC). Methods: 1. Cytotoxicity of allicin to NCI-H460 (1) the inhibitory effect of allicin on the proliferation of NCI-H460 cells was detected by MTT assay; (2) the apoptosis rate of NCI-H460 cells was detected by flow cytometry; (3) the activity of apoptosis-related factor Caspase-3 was detected by colorimetry; 2. Antitumor effect of allicin on Lewis lung cancer mice (1) culture of Lewis lung cancer cell line, (2) establishment of transplanted tumor model of Lewis lung cancer in mice, (3) killing mice, calculation of tumor inhibition rate, spleen index and thymus index, (4) the morphology of the cells was observed after HE staining. Results: 1. After treated with allicin for 24 h, the morphology of NCI-H460 cells was observed under microscope. After treated with garlicine, the shape of NCI-H460 cells was irregular, the cells shrank, the size of the cells was different, and the degree of cell fragmentation increased with the increase of dosages. The number of cells gradually decreased. 2. When NCI-H460 cells were treated with garlicine for 24 h, the proliferation of NCI-H460 cells was significantly inhibited (p0.05, p0.01), and the IC50 was 5.79 鹵0.11 渭 mol / L, compared with the control group (p0.05, p0.01), and the IC50 was 5.79 鹵0.11 渭 mol / L, and the IC50 was 5.79 鹵0.11 渭 mol / L. It is suggested that allicin can effectively inhibit the proliferation of lung cancer NCI-H460 cells. 3. With the increase of the concentration of allicin, the apoptosis rate of the treated group increased gradually, and the difference was significant compared with that of the saline group (p0.01). At the highest concentration of 16 渭 mol / L, the apoptosis rate reached 46.2 鹵3.46%. These results suggest that Lycorine can induce apoptosis of NCI-H460 cells in a dose-dependent manner. 4. With the increase of the concentration of allicin, the activity of Caspase-3 increased gradually. Compared with the control group, the activity of Caspase-3 was significantly increased in the garlicine treatment group (p0.01). The results showed that allicin could enhance the activity of Caspase-3 in lung cancer NCI-H460 cells and induce the apoptosis of NCI-H460 cells. 5. Compared with normal saline group, Lycorine had inhibitory effect on tumor growth of Lewis lung cancer mice in low, middle and high dose groups, among which the high dose group (40 mg/kg) had the most significant effect. The tumor inhibition rate was 56.8% (p0.01). The results of HE staining in solid tumor tissue of tumor-bearing mice showed that the tumor cells in the saline group grew uniformly and closely with less nuclear condensation and no necrosis or punctate necrosis. A large number of nuclear pyknosis, nuclear fragmentation, decrease in the number of tumor cells and loose arrangement were observed in various dose groups and cyclophosphamide groups, especially in the high dose group (40 mg/kg), and the distribution of tumor cells was obvious. There was a marked decrease in the number. Conclusion: garlicine can effectively inhibit the proliferation and promote the apoptosis of NCI-H460 cells in vitro. The mechanism may be related to the activation of Caspase-3 expression and the induction of apoptosis of tumor cells. Allicin can also inhibit the growth of Lewis lung cancer cells in vivo.
【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R734.2
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