多糖GF对荷瘤小鼠H_(22)细胞P53、Rb基因以及E2F转录因子的影响
摘要
树舌多糖GF是从中药树舌干燥子实体中提取的一种有效成分,已有一些研究表明,中药树舌多糖GF在抗肿瘤、提高机体免疫力方面起重要作用。因此,本实验研究树舌多糖GF对荷瘤小鼠H22细胞P53基因、Rb基因以及E2F转录因子表达的影响,旨在探讨树舌多糖GF的抗肿瘤机理,丰富树舌多糖的作用机制。
目的：
为探明中药树舌多糖GF对肝癌小鼠H22细胞P53基因、Rb基因以及E2F转录因子的影响。从分子生物学水平研究中药树舌多糖的抗肿瘤作用机制以及小鼠用药后,肝癌细胞中P53基因、Rb基因以及E2F转录因子表达的变化。
方法：
实验共分为四组进行。正常对照组为正常小鼠,每天给予生理盐水0.1ml/只。阴性对照组为肝癌H22荷瘤小鼠,每天给予生理盐水0.1ml/只阳性对照组为肝癌H22荷瘤小鼠,每天给予环磷酰胺3.0mg/kg·d-1。树舌多糖组为肝癌H22荷瘤小鼠,每天给予树舌多糖GF注射液50mg/kg·d-1。待阴性对照组的小鼠肿瘤长到1g左右,将各组小鼠全部处死。取出各组完整的肿瘤组织,称重。正常组取肝脏。用ELISA法检测P53基因、Rb基因以及E2F转录因子表达的情况并进行分析。
结果：
1.树舌多糖GF组中P53基因的蛋白表达量均显著高于阴性对照组(P<0.01)。树舌多糖GF组与环磷酰胺组P53基因的蛋白表达量无显著差异
2.树舌多糖GF组中Rb基因的蛋白表达量均显著高于阴性对照组(P<0.01)。树舌多糖GF组与环磷酰胺组Rb基因的蛋白表达量无显著差异。
3.树舌多糖GF组中E2F转录因子表达量均显著低于阴性对照组(P<0.01)。树舌多糖GF组与环磷酰胺组E2F转录因子的表达量无显著差异。
4.树舌多糖组、环磷酰胺组P53基因、Rb基因以及E2F转录因子的表达量无显著差异。
结论：
1.树舌多糖GF可使肝癌小鼠H22细胞P53基因蛋白的表达上调。
2.树舌多糖GF可使肝癌小鼠H22细胞Rb基因蛋白的表达上调。
3.树舌多糖GF可使肝癌小鼠H22细胞E2F转录因子的表达下调。
GAPSGF is a kind of effective component extracted from ganoderma appanatum. There is some research shows that GAPSGF has important effect on antitumor,enhancing immunity. So this experiment is studying the effect on P53、Rb and E2F in H22 tumor cell with GAPSGF. Aim is to study the antitumor mechanism of GAPSGF and its mechanism.
Objective:
From the level of molecular biology to study the antitumor mechanism of GAPSGF and the effect on RB、P53 and E2F in H22 tumor cell with GAPSGF.
Methods:
There are four groups in the experiment. The normal control group was given normal saline. The negative group was given normal saline too. The positive control group was given cyclophosphamide according to 30mg/kg·d-1, GAPSGF group was given the medicine according to 50mg/kg·d-1. When the negative control group mice tumor achieves above 1g, then execute the mice the next day. Then peel entire tumor tissue of every group, weight, calculate tumor inhibit rate. Enzyme-linked immunoassay method was used to determine the RB、P53 and E2F gene expression level before and after treatment.
Results:
1. significantly enhancing-the P53 expression compared with those in the control group with p<0.01
2. significantly enhancing the Rb expression compared with those in the control group with p<0.01
3. significantly reducing the E2F expression compared with those in the control group with p<0.01
4. There was no significant difference between GAPSGF and CTX.
Conclusions:
1. GAPSGF could increase the expression of P53 in hepatoma H22 tumor tissue.
2.GAPSGF could increase the expression of RB in hepatoma H22 tumor tissue.
3.GAPSGF could reduce the expression of E2F in hepatoma H22 tumor tissue.
目的：
为探明中药树舌多糖GF对肝癌小鼠H22细胞P53基因、Rb基因以及E2F转录因子的影响。从分子生物学水平研究中药树舌多糖的抗肿瘤作用机制以及小鼠用药后,肝癌细胞中P53基因、Rb基因以及E2F转录因子表达的变化。
方法：
实验共分为四组进行。正常对照组为正常小鼠,每天给予生理盐水0.1ml/只。阴性对照组为肝癌H22荷瘤小鼠,每天给予生理盐水0.1ml/只阳性对照组为肝癌H22荷瘤小鼠,每天给予环磷酰胺3.0mg/kg·d-1。树舌多糖组为肝癌H22荷瘤小鼠,每天给予树舌多糖GF注射液50mg/kg·d-1。待阴性对照组的小鼠肿瘤长到1g左右,将各组小鼠全部处死。取出各组完整的肿瘤组织,称重。正常组取肝脏。用ELISA法检测P53基因、Rb基因以及E2F转录因子表达的情况并进行分析。
结果：
1.树舌多糖GF组中P53基因的蛋白表达量均显著高于阴性对照组(P<0.01)。树舌多糖GF组与环磷酰胺组P53基因的蛋白表达量无显著差异
2.树舌多糖GF组中Rb基因的蛋白表达量均显著高于阴性对照组(P<0.01)。树舌多糖GF组与环磷酰胺组Rb基因的蛋白表达量无显著差异。
3.树舌多糖GF组中E2F转录因子表达量均显著低于阴性对照组(P<0.01)。树舌多糖GF组与环磷酰胺组E2F转录因子的表达量无显著差异。
4.树舌多糖组、环磷酰胺组P53基因、Rb基因以及E2F转录因子的表达量无显著差异。
结论：
1.树舌多糖GF可使肝癌小鼠H22细胞P53基因蛋白的表达上调。
2.树舌多糖GF可使肝癌小鼠H22细胞Rb基因蛋白的表达上调。
3.树舌多糖GF可使肝癌小鼠H22细胞E2F转录因子的表达下调。
GAPSGF is a kind of effective component extracted from ganoderma appanatum. There is some research shows that GAPSGF has important effect on antitumor,enhancing immunity. So this experiment is studying the effect on P53、Rb and E2F in H22 tumor cell with GAPSGF. Aim is to study the antitumor mechanism of GAPSGF and its mechanism.
Objective:
From the level of molecular biology to study the antitumor mechanism of GAPSGF and the effect on RB、P53 and E2F in H22 tumor cell with GAPSGF.
Methods:
There are four groups in the experiment. The normal control group was given normal saline. The negative group was given normal saline too. The positive control group was given cyclophosphamide according to 30mg/kg·d-1, GAPSGF group was given the medicine according to 50mg/kg·d-1. When the negative control group mice tumor achieves above 1g, then execute the mice the next day. Then peel entire tumor tissue of every group, weight, calculate tumor inhibit rate. Enzyme-linked immunoassay method was used to determine the RB、P53 and E2F gene expression level before and after treatment.
Results:
1. significantly enhancing-the P53 expression compared with those in the control group with p<0.01
2. significantly enhancing the Rb expression compared with those in the control group with p<0.01
3. significantly reducing the E2F expression compared with those in the control group with p<0.01
4. There was no significant difference between GAPSGF and CTX.
Conclusions:
1. GAPSGF could increase the expression of P53 in hepatoma H22 tumor tissue.
2.GAPSGF could increase the expression of RB in hepatoma H22 tumor tissue.
3.GAPSGF could reduce the expression of E2F in hepatoma H22 tumor tissue.
引文
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