自拟方抑癌汤对人肺癌NCI-H446细胞增殖抑制作用的体外实验研究
|
摘要
目的:本课题应用细胞培养技术,通过自拟方抑癌汤对人肺癌NCI-H446细胞株进行干扰研究,探讨其增殖抑制作用。旨在从苗药中药组方中找到抑制肺癌细胞增殖的新方法,为治疗肺癌提供一种新的治疗思路。
方法:将NCI-H446细胞进行培养,绘制NCI-H446细胞生长曲线,取处在对数生长期的NCI-H446细胞进行实验。首先确定抑癌汤的作用浓度范围,将抑癌汤分为1g/ml、100mg/ml、10 mg/ml、1 mg/ml、100ug/ml、10 ug/ml、1 ug/ml、0.1ug/ml共8个浓度梯度,对照组为只加等量的细胞培养液,总共9组。每一组各设4个复孔,使用MTT法测定细胞的存活率。取细胞的存活率为50%时的药物浓度范围做为抑癌汤的给药参考浓度。再将抑癌汤在20ug/ml、40ug/ml、60ug/ml、80ug/ml、100ug/ml五个浓度和在24小时、48小时、72小时这三个观测时间点上与空白组(培养基对照组)进行比较观察。最后取抑癌汤60ug/ml的浓度做为抑癌汤的给药浓度,与西药组;抑癌汤加西药组(60ug/ml浓度组并加用西药)和以只加培养液的空白对照组,共四个大组(抑癌汤组、西药组、抑癌汤加西药组、空白组),每一组各设4个复孔,进行对照研究,使用MTT法测定细胞的抑制率,以上实验均重复三次。
结果:
1:绘制细胞生长曲线,通过倒置显微镜下进行细胞计数,得到细胞倍增时间为26小时。
2:抑癌汤对NCI-H446细胞毒实验得到细胞存活率50%时的药物浓度范围在1-100ug/ml之间,根据该浓度范围可以为细胞增殖抑制实验提供给药药物参考浓度。
3:细胞增殖抑制实验:抑癌汤在20ug/ml、40ug/ml、60ug/ml、80ug/ml、100ug/ml浓度和在24小时、48小时、72小时这三个观测时间点与空白组(培养基对照组)比较均有统计学差异。抑癌汤组三次实验经过统计分析得到:P=0.492,即抑癌汤组三次重复实验无统计学差异,说明实验具有可重复性。结果显示不同浓度抑癌汤作用48h对NCI-H446细胞的增殖具有明显的抑制作用,随着作用时间的延长抑制作用逐渐减弱,且随着给药剂量的增大,抑制作用增强。抑癌汤在24小时、48小时、72小时的抑制率分别达到53.47%、54.47%和10.45%。
4:四个给药组之间的比较:抑癌汤组选用的中间浓度(60ug/ml)。抑癌汤加西药组与抑癌汤组;抑癌汤加西药组与西药组;抑癌汤组与西药组;抑癌汤组与空白组;抑癌汤加西药组与空白组;西药组和空白组的细胞抑制率有统计学差异。
通过给药组间均数两两比较:抑癌汤加西药组优于抑癌汤组;抑癌汤加西药组优于西药组;西药组优于抑癌汤组;抑癌汤组优于空白组;抑癌汤加西药组优于空白组;西药组优于空白组并有统计学差异。
结论:
1:人小细胞肺癌NCI-H446细胞倍增时间为26小时,增殖快。
2:结果显示不同浓度抑癌汤作用48h对NCI-H446细胞的增殖具有明显的抑制作用,随着作用时间的延长抑制作用逐渐减弱。
3:抑癌汤对NCI-H446细胞随着给药剂量的增大,抑制作用增强。
4:抑癌汤加西药组优于抑癌汤组;抑癌汤加西药组优于西药组;西药组优于抑癌汤组。
抑癌汤在体外对人小细胞肺癌NCI-H446细胞具有抑制作用,通过理气、活血止痛、燥湿祛痰、清热解毒的方法抑制癌细胞增殖。具有潜在药用价值,值得进一步深入研究。
Objective: The topic cell culture technology, Since the adoption of the proposed side suppressor soup on human lung cancer cell line NCI-H446 interference, explore its proliferation. From seedling to the Chinese Medicines board prescription drugs found lung cancer cell proliferation inhibition new method for the treatment of lung cancer with a new treatment ideas.
Methods: NCI-H446 cells were cultured in plotting NCI-H446 cells growth curve, from the log at the NCI-H446 cells for experiments. First determine the role of tumor suppressor soup concentration of tumor suppressor soup into 1g/ml,100mg/ml.10mg/ml,1mg/ml,100ug/ml,10ug/ml, 1ug/ml, 0.1ug/ml a total of eight concentration gradient, the control group only increase contour cell culture media, a total of nine. Each group set up four Minute hole, MTT assay was used to measure cell survival. Cells from the 50% survival rate for the drug concentration as a tumor suppressor soup administrationb reference concentration. In further unrest 20ug/ml,40ug/ml,60ug/ml,80ug/ml suppressor, 100ug/ml 5 concentration and 24 hours, 48 hours, This 72-hour observation time three point blank (media control group) were compared observation. For the final soup 60ug/ml suppressor as the concentration of tumor suppressor soup administration concentration, and the Western group. suppressor Tonga medicine group (60ug/ml concentrations and increases with western medicine) and the only culture fluid of the control group. A total of four larger groups (suppressor of group, medicine group, the tumor suppressor Tonga medicine group, the control group), a group set up four of the Minute hole, Comparative study, the MTT assay was used to measure the cell growth rate, the above experiments were repeated three times.
Results:1 : Drawing cell growth curve inverted microscope for cell counting, cell doubling time was 26 hours. 2 : NCI-H446 tumor suppressor soup of cytotoxic cell experiments at the 50% survival rate of drug concentration in the range of 1-10 0ug/ml between, according to the concentration range for the inhibition of cell proliferation with reference to the concentration of drug administration. 3 : inhibition of cell proliferation : the tumor suppressor 20ug/ml,40ug/ml,60ug/ml soup, 80ug/ml,100ug/ml concentration and 24 hours, 48 hours, This 72-hour observation time three point blank Group (media control group) were compared statistically significant difference. Suppressor soup three experimental groups after statistical analysis : P=0.492. suppressor soup that is repeated three experimental groups no significant difference on experiments with repeatability. The results showed that different concentrations of tumor suppressor role of soup for 48 hours NCI-H446 cells was inhibited, With the duration of effect gradually weakened, and with the dosage increased, the inhibition. Suppressor soup in 24 hours, 48 hours, 72 hours of inhibition rate reached 53.47%, respectively, 54.47% and 10.45%.4 : four administration of comparisons between groups : Group selected tumor suppressor soup in the middle concentration (60ug/ml). Suppressor Tonga medicine group and tumor suppressor soup; Suppressor Tonga western medicine and western medicine; Suppressor soup and western medicine; suppressor soup group and control group; suppressor Tonga medicine group and control group; Western and blank cells inhibition rate was statistically different. Through the administration of several groups were compared on February 2 : suppressor Tonga medicine group than suppressor soup; Suppressor Tonga better than Western medicine group; WM group than suppressor soup; suppressor soup is better than the blank group; suppressor Tonga medicine group is better than the blank group; Western medicine is better than the blank group and the significant difference.
Conclusion: 1 : human small cell lung cancer NCI-H446 cells doubling time of 26 hours, rapid proliferation. 2 : The results showed that different concentrations of tumor suppressor role of soup for 48 hours NCI-H446 cells was inhibited, With the duration of effect gradually diminish. 3 : soup of NCI-H446 tumor suppressor cells with the dosage increased, the inhibition. 4 : Tonga medicine group suppressor tumor suppressor than soup; Suppressor Tonga better than Western medicine group; WM group than suppressor Soup. Suppressor unrest in vitro human small cell lung cancer NCI-H446 cells were inhibited by Shugan Jimmy gas, Huoxuezhitong, and removing expectorant, Qingrejiedu inhibition of cell proliferation. Have potential medicinal value, it is worth further study.
方法:将NCI-H446细胞进行培养,绘制NCI-H446细胞生长曲线,取处在对数生长期的NCI-H446细胞进行实验。首先确定抑癌汤的作用浓度范围,将抑癌汤分为1g/ml、100mg/ml、10 mg/ml、1 mg/ml、100ug/ml、10 ug/ml、1 ug/ml、0.1ug/ml共8个浓度梯度,对照组为只加等量的细胞培养液,总共9组。每一组各设4个复孔,使用MTT法测定细胞的存活率。取细胞的存活率为50%时的药物浓度范围做为抑癌汤的给药参考浓度。再将抑癌汤在20ug/ml、40ug/ml、60ug/ml、80ug/ml、100ug/ml五个浓度和在24小时、48小时、72小时这三个观测时间点上与空白组(培养基对照组)进行比较观察。最后取抑癌汤60ug/ml的浓度做为抑癌汤的给药浓度,与西药组;抑癌汤加西药组(60ug/ml浓度组并加用西药)和以只加培养液的空白对照组,共四个大组(抑癌汤组、西药组、抑癌汤加西药组、空白组),每一组各设4个复孔,进行对照研究,使用MTT法测定细胞的抑制率,以上实验均重复三次。
结果:
1:绘制细胞生长曲线,通过倒置显微镜下进行细胞计数,得到细胞倍增时间为26小时。
2:抑癌汤对NCI-H446细胞毒实验得到细胞存活率50%时的药物浓度范围在1-100ug/ml之间,根据该浓度范围可以为细胞增殖抑制实验提供给药药物参考浓度。
3:细胞增殖抑制实验:抑癌汤在20ug/ml、40ug/ml、60ug/ml、80ug/ml、100ug/ml浓度和在24小时、48小时、72小时这三个观测时间点与空白组(培养基对照组)比较均有统计学差异。抑癌汤组三次实验经过统计分析得到:P=0.492,即抑癌汤组三次重复实验无统计学差异,说明实验具有可重复性。结果显示不同浓度抑癌汤作用48h对NCI-H446细胞的增殖具有明显的抑制作用,随着作用时间的延长抑制作用逐渐减弱,且随着给药剂量的增大,抑制作用增强。抑癌汤在24小时、48小时、72小时的抑制率分别达到53.47%、54.47%和10.45%。
4:四个给药组之间的比较:抑癌汤组选用的中间浓度(60ug/ml)。抑癌汤加西药组与抑癌汤组;抑癌汤加西药组与西药组;抑癌汤组与西药组;抑癌汤组与空白组;抑癌汤加西药组与空白组;西药组和空白组的细胞抑制率有统计学差异。
通过给药组间均数两两比较:抑癌汤加西药组优于抑癌汤组;抑癌汤加西药组优于西药组;西药组优于抑癌汤组;抑癌汤组优于空白组;抑癌汤加西药组优于空白组;西药组优于空白组并有统计学差异。
结论:
1:人小细胞肺癌NCI-H446细胞倍增时间为26小时,增殖快。
2:结果显示不同浓度抑癌汤作用48h对NCI-H446细胞的增殖具有明显的抑制作用,随着作用时间的延长抑制作用逐渐减弱。
3:抑癌汤对NCI-H446细胞随着给药剂量的增大,抑制作用增强。
4:抑癌汤加西药组优于抑癌汤组;抑癌汤加西药组优于西药组;西药组优于抑癌汤组。
抑癌汤在体外对人小细胞肺癌NCI-H446细胞具有抑制作用,通过理气、活血止痛、燥湿祛痰、清热解毒的方法抑制癌细胞增殖。具有潜在药用价值,值得进一步深入研究。
Objective: The topic cell culture technology, Since the adoption of the proposed side suppressor soup on human lung cancer cell line NCI-H446 interference, explore its proliferation. From seedling to the Chinese Medicines board prescription drugs found lung cancer cell proliferation inhibition new method for the treatment of lung cancer with a new treatment ideas.
Methods: NCI-H446 cells were cultured in plotting NCI-H446 cells growth curve, from the log at the NCI-H446 cells for experiments. First determine the role of tumor suppressor soup concentration of tumor suppressor soup into 1g/ml,100mg/ml.10mg/ml,1mg/ml,100ug/ml,10ug/ml, 1ug/ml, 0.1ug/ml a total of eight concentration gradient, the control group only increase contour cell culture media, a total of nine. Each group set up four Minute hole, MTT assay was used to measure cell survival. Cells from the 50% survival rate for the drug concentration as a tumor suppressor soup administrationb reference concentration. In further unrest 20ug/ml,40ug/ml,60ug/ml,80ug/ml suppressor, 100ug/ml 5 concentration and 24 hours, 48 hours, This 72-hour observation time three point blank (media control group) were compared observation. For the final soup 60ug/ml suppressor as the concentration of tumor suppressor soup administration concentration, and the Western group. suppressor Tonga medicine group (60ug/ml concentrations and increases with western medicine) and the only culture fluid of the control group. A total of four larger groups (suppressor of group, medicine group, the tumor suppressor Tonga medicine group, the control group), a group set up four of the Minute hole, Comparative study, the MTT assay was used to measure the cell growth rate, the above experiments were repeated three times.
Results:1 : Drawing cell growth curve inverted microscope for cell counting, cell doubling time was 26 hours. 2 : NCI-H446 tumor suppressor soup of cytotoxic cell experiments at the 50% survival rate of drug concentration in the range of 1-10 0ug/ml between, according to the concentration range for the inhibition of cell proliferation with reference to the concentration of drug administration. 3 : inhibition of cell proliferation : the tumor suppressor 20ug/ml,40ug/ml,60ug/ml soup, 80ug/ml,100ug/ml concentration and 24 hours, 48 hours, This 72-hour observation time three point blank Group (media control group) were compared statistically significant difference. Suppressor soup three experimental groups after statistical analysis : P=0.492. suppressor soup that is repeated three experimental groups no significant difference on experiments with repeatability. The results showed that different concentrations of tumor suppressor role of soup for 48 hours NCI-H446 cells was inhibited, With the duration of effect gradually weakened, and with the dosage increased, the inhibition. Suppressor soup in 24 hours, 48 hours, 72 hours of inhibition rate reached 53.47%, respectively, 54.47% and 10.45%.4 : four administration of comparisons between groups : Group selected tumor suppressor soup in the middle concentration (60ug/ml). Suppressor Tonga medicine group and tumor suppressor soup; Suppressor Tonga western medicine and western medicine; Suppressor soup and western medicine; suppressor soup group and control group; suppressor Tonga medicine group and control group; Western and blank cells inhibition rate was statistically different. Through the administration of several groups were compared on February 2 : suppressor Tonga medicine group than suppressor soup; Suppressor Tonga better than Western medicine group; WM group than suppressor soup; suppressor soup is better than the blank group; suppressor Tonga medicine group is better than the blank group; Western medicine is better than the blank group and the significant difference.
Conclusion: 1 : human small cell lung cancer NCI-H446 cells doubling time of 26 hours, rapid proliferation. 2 : The results showed that different concentrations of tumor suppressor role of soup for 48 hours NCI-H446 cells was inhibited, With the duration of effect gradually diminish. 3 : soup of NCI-H446 tumor suppressor cells with the dosage increased, the inhibition. 4 : Tonga medicine group suppressor tumor suppressor than soup; Suppressor Tonga better than Western medicine group; WM group than suppressor Soup. Suppressor unrest in vitro human small cell lung cancer NCI-H446 cells were inhibited by Shugan Jimmy gas, Huoxuezhitong, and removing expectorant, Qingrejiedu inhibition of cell proliferation. Have potential medicinal value, it is worth further study.
引文
[1] 章静波.组织和细胞培养技术.研究生规划教材.北京:人民卫生出版社,2002.111～112
[2] 刘刚.Excel 在统计分析中的应用. 北京:人民卫生出版社,2002.170～174
[3] 熊敏,吴一龙.现代肺癌病理与临床.北京:科学出版社.医学出版分社,2003.86～87
[4] 徐振晔,杨宇飞.肺癌中西医综合治疗.北京:人民卫生出版社,2002.1
[5] 王吉耀.内科学.供 8 年制及 7 年制临床医学等专业用.北京:人民卫生出版社,2005.112
[6] 徐振晔,杨宇飞.肺癌中西医综合治疗.北京:人民卫生出版社,2002.15
[7] 陈锐深.现代中医肿瘤学. 北京:人民卫生出版社,2003.1
[8] 王永炎.中医内科学.普通高等教育中医药类规划教材.上海:上海科学技术出版社,1997.91
[9] 辛 海.肺癌的中医临床治疗进展.Beijing Journal of TCM,2003;22(6):50
[10] 陈锐深.现代中医肿瘤学.北京:人民卫生出版社 2003:3
[11] 奇玲,罗达尚.中国少数民族传统医药大系.内蒙古:内蒙古科学技术出版社,2000.1
[12] 周厚琼,冉懋雄.中国贵州苗药产业的特色与蓬勃发展
[13] 奇玲,罗达尚.中国少数民族传统医药大系.内蒙古:内蒙古科学技术出版社,2000.539～584
[14] 国家药典委员会.中华人民共和国药典.一部.北京:人民卫生出版社,1990:266
[15] 汪毅.中国苗族药物彩色图集.贵州:贵州科学技术出版社.2002:28
[16] 何洪源.中国粗榧果实和枝叶中三尖杉酯碱和高三尖杉酯碱分离方法的研究.中草药,2001;32(3):201-202
[17] 任仁安.中药鉴定学[M].上海:上海科学技术出版社,1986:491
[18] 蔡楚伧,钱秀丽,姜达卫.白花蛇舌草化学成分研究Ⅰ[J].药学学报,1964,11(2):809
[19] 吕华冲,何军.白花蛇舌草化学成分的研究[J].广东医药学院学报,1994,10(2):125
[20] 方岩雄,张永成,陈敏敏,等.抗肿瘤药物白花蛇舌草及其活性成分[J].中成药,2004,26(7):577
[21] 于 新,杜志强,陈悦娇,等.白花蛇舌草提取物抗氧化作用的研究[J].食品与发酵工业,2001,28(3):10
[22] 周建波,龙斯华,黄存礼,等.白花蛇舌草的微量元素分析[J].中国中医药杂志,1990,15(12):36
[23] 雷载权.中药学.上海:上海科学技术出版社,1995:84
[24] 张伟平,尹秀丽.白花蛇舌草的研究[J].哈尔滨医药,2000,20(2):71
[25] 陈仁寿.国家药典中药实用手册.南京:江苏科学技术出版社,2004.126
[26] 季宇彬.抗癌中药药理与应用.哈尔滨:黑龙江科学技术出版社,1999.443
[27]章永红.抗癌抑癌汤大全.南京:江苏科学技术出版社,2000,6
[28] 刘嘉湘.施志明.徐振哗,等.金复康口服液治疗非小细胞肺癌的临床观察[J].中医杂志,1997,38(12):72
[29] 刘嘉湘,潘敏求,黎月恒,等.金复康口服液治疗原发性非小细胞肺癌临床研究.肿瘤,2001,21(6):463
[30] 孙建立.中医药治疗肺癌的机理研究概况. 上海中医药大学学报,2001,15(1):61
[1] R.I.弗雷谢尼[英] 编著,章静波,徐存拴,等译.动物细胞培养——基本技术指南(第四版).北京:科学出版社,2004,3
[2] 熊敏,吴一龙.现代肺癌病理与临床.第一版.北京:科技出版社,2003.23
[3] 刘 茜 , 苏 勤 , 刘 节 .Neuron specific enolase,P53 蛋 白 和proliferating-cell nuclear antigen 在肺癌组织中的表达及意义.第四军医大学学报,1999;(20):246
[4] 郭广松,刘建军,徐世杰.VEGF、PCNA 在肺癌组织中的表达及其意义.武汉大学学报.医学版,2001;(22):12
[5] 马晓春,张玲,马红.肺癌血管内皮生长因子表达及其与血管新生和肿瘤细胞增殖关系的研究.中华内科杂志,2001;(40):32
[6] 黄群.扶正解毒汤治疗晚期肺癌 90 例临床观察.江西中医药,2004;(7):32
[7] 熊玮,沈寒放.对活检肺癌组织行原代细胞培养及药敏试验的研究.中华结核和呼吸杂志,1997;(20):2
[8] 谢启梅口述.谢军荣整理.龙葵在治疗肿瘤中的应用.江西中医药,1996;(27):52
[9] Goldman and bennett[美] 总主编,王贤才主译.西氏内科学.第 21 版.北京:世界图书出版公司,2003,45
[10] 连华,周锦明.中医药治疗肺癌的体会.中国临床医生,1999;(27):46
[11] 徐振华,杨宇飞.肺癌中西医结合治疗.第一版 .北京:人民卫生出版社,2002,56
[12] 殷国荣,王斌全,杨建一.医学科研方法与论文写作.北京:科学出版社,2002,45
[13] 张东方,黄炜,黄济群.齐墩果酸抗人肺癌细胞增殖、侵袭和诱导细胞凋亡的研究.肺癌防治研究,2003;(30):3
[2] 刘刚.Excel 在统计分析中的应用. 北京:人民卫生出版社,2002.170～174
[3] 熊敏,吴一龙.现代肺癌病理与临床.北京:科学出版社.医学出版分社,2003.86～87
[4] 徐振晔,杨宇飞.肺癌中西医综合治疗.北京:人民卫生出版社,2002.1
[5] 王吉耀.内科学.供 8 年制及 7 年制临床医学等专业用.北京:人民卫生出版社,2005.112
[6] 徐振晔,杨宇飞.肺癌中西医综合治疗.北京:人民卫生出版社,2002.15
[7] 陈锐深.现代中医肿瘤学. 北京:人民卫生出版社,2003.1
[8] 王永炎.中医内科学.普通高等教育中医药类规划教材.上海:上海科学技术出版社,1997.91
[9] 辛 海.肺癌的中医临床治疗进展.Beijing Journal of TCM,2003;22(6):50
[10] 陈锐深.现代中医肿瘤学.北京:人民卫生出版社 2003:3
[11] 奇玲,罗达尚.中国少数民族传统医药大系.内蒙古:内蒙古科学技术出版社,2000.1
[12] 周厚琼,冉懋雄.中国贵州苗药产业的特色与蓬勃发展
[13] 奇玲,罗达尚.中国少数民族传统医药大系.内蒙古:内蒙古科学技术出版社,2000.539～584
[14] 国家药典委员会.中华人民共和国药典.一部.北京:人民卫生出版社,1990:266
[15] 汪毅.中国苗族药物彩色图集.贵州:贵州科学技术出版社.2002:28
[16] 何洪源.中国粗榧果实和枝叶中三尖杉酯碱和高三尖杉酯碱分离方法的研究.中草药,2001;32(3):201-202
[17] 任仁安.中药鉴定学[M].上海:上海科学技术出版社,1986:491
[18] 蔡楚伧,钱秀丽,姜达卫.白花蛇舌草化学成分研究Ⅰ[J].药学学报,1964,11(2):809
[19] 吕华冲,何军.白花蛇舌草化学成分的研究[J].广东医药学院学报,1994,10(2):125
[20] 方岩雄,张永成,陈敏敏,等.抗肿瘤药物白花蛇舌草及其活性成分[J].中成药,2004,26(7):577
[21] 于 新,杜志强,陈悦娇,等.白花蛇舌草提取物抗氧化作用的研究[J].食品与发酵工业,2001,28(3):10
[22] 周建波,龙斯华,黄存礼,等.白花蛇舌草的微量元素分析[J].中国中医药杂志,1990,15(12):36
[23] 雷载权.中药学.上海:上海科学技术出版社,1995:84
[24] 张伟平,尹秀丽.白花蛇舌草的研究[J].哈尔滨医药,2000,20(2):71
[25] 陈仁寿.国家药典中药实用手册.南京:江苏科学技术出版社,2004.126
[26] 季宇彬.抗癌中药药理与应用.哈尔滨:黑龙江科学技术出版社,1999.443
[27]章永红.抗癌抑癌汤大全.南京:江苏科学技术出版社,2000,6
[28] 刘嘉湘.施志明.徐振哗,等.金复康口服液治疗非小细胞肺癌的临床观察[J].中医杂志,1997,38(12):72
[29] 刘嘉湘,潘敏求,黎月恒,等.金复康口服液治疗原发性非小细胞肺癌临床研究.肿瘤,2001,21(6):463
[30] 孙建立.中医药治疗肺癌的机理研究概况. 上海中医药大学学报,2001,15(1):61
[1] R.I.弗雷谢尼[英] 编著,章静波,徐存拴,等译.动物细胞培养——基本技术指南(第四版).北京:科学出版社,2004,3
[2] 熊敏,吴一龙.现代肺癌病理与临床.第一版.北京:科技出版社,2003.23
[3] 刘 茜 , 苏 勤 , 刘 节 .Neuron specific enolase,P53 蛋 白 和proliferating-cell nuclear antigen 在肺癌组织中的表达及意义.第四军医大学学报,1999;(20):246
[4] 郭广松,刘建军,徐世杰.VEGF、PCNA 在肺癌组织中的表达及其意义.武汉大学学报.医学版,2001;(22):12
[5] 马晓春,张玲,马红.肺癌血管内皮生长因子表达及其与血管新生和肿瘤细胞增殖关系的研究.中华内科杂志,2001;(40):32
[6] 黄群.扶正解毒汤治疗晚期肺癌 90 例临床观察.江西中医药,2004;(7):32
[7] 熊玮,沈寒放.对活检肺癌组织行原代细胞培养及药敏试验的研究.中华结核和呼吸杂志,1997;(20):2
[8] 谢启梅口述.谢军荣整理.龙葵在治疗肿瘤中的应用.江西中医药,1996;(27):52
[9] Goldman and bennett[美] 总主编,王贤才主译.西氏内科学.第 21 版.北京:世界图书出版公司,2003,45
[10] 连华,周锦明.中医药治疗肺癌的体会.中国临床医生,1999;(27):46
[11] 徐振华,杨宇飞.肺癌中西医结合治疗.第一版 .北京:人民卫生出版社,2002,56
[12] 殷国荣,王斌全,杨建一.医学科研方法与论文写作.北京:科学出版社,2002,45
[13] 张东方,黄炜,黄济群.齐墩果酸抗人肺癌细胞增殖、侵袭和诱导细胞凋亡的研究.肺癌防治研究,2003;(30):3