藓化饮对DMBA诱导的金黄地鼠颊黏膜癌前病变逆转作用的实验研究
|
摘要
目的:口腔黏膜癌前病变是一种癌前状态,表现为口腔黏膜上皮细胞出现不同程度的不典型增生,但尚未发展成癌。新的WHO分类中指出癌前病变的最新定义为:“转变为鳞状细胞癌的可能性增加的发生改变的上皮。这种改变的上皮出现各种传统上称为上皮异常增生的细胞学和组织结构的变化”。目前公认的癌前病变和癌前状态包括白斑、红斑、扁平苔藓和口腔黏膜下纤维性变等。癌前病变如果不能及时治疗而任其发展,最终会发展成癌,严重影响治疗效果和预后,也给患者的生理及心理造成巨大影响。如何对口腔黏膜癌前病变进行治疗,阻断其向口腔癌发展,已经成为当前研究的热点。目前针对口腔黏膜癌前病变的治疗方法较多,主要有西药口服、全身及局部应用激素和免疫抑制剂、早期手术切除及中药治疗等。但应用西药疗效不稳定,激素及免疫抑制剂初期起效虽快但作用不持久且副作用大,而以往的中药治疗多结合局部激素应用,单纯应用中药治疗癌前病变的报道较少,缺乏足够的实验研究来证实中药的治疗效果。针对口腔黏膜癌前病变的特点,中医中药在治疗口腔黏膜癌前病变方面具有明显的优势:中药标本同治、疗效稳固;口服汤剂或中成药制剂,服用方法简便,患者的依从性好,易于接受;与西药相比副作用小等。鉴于以上的优势,中药在治疗癌前病变方面越来越受到人们的重视。本实验采用中药藓化饮对实验动物口腔黏膜癌前病变进行治疗干预,并通过流式细胞术和免疫组化方法观察治疗前后P53和COX-2表达情况,分析和探讨藓化饮对口腔黏膜癌前病变的逆转作用。为今后临床应用提供实验理论依据。
方法:
1.口腔黏膜癌前病变动物模型制备及分组
取叙利亚金黄地鼠(SPF级)55只,6～8周龄,体长约10cm,体重90～110g,雌雄兼备,由北京维通利华实验动物技术有限公司提供,生产许可证编号:SCXK(京)2007-0001。按照完全随机化原则分为四组:空白对照组5只,癌前病变对照组10只,生理盐水治疗对照组10只,藓化饮治疗组30只。
空白对照组每日正常饲养,不做处理,直至实验结束,与其余组一同处死并取材。
癌前病变对照组、生理盐水治疗对照组、藓化饮治疗组均以0.5%DMBA涂抹双侧颊黏膜,每周三次,持续六周。
2.中药藓化饮煎剂的制备
先向组方中药中加入约8～10倍的水,浸泡40分钟～1小时,用武火加热20～30分钟至煮沸,保持约10分钟,再以文火煎约10分钟后,过滤,取汁;再加入相当于剩余滤渣量6倍的水,用武火加热20～30分钟至煮沸,用文火煎约10分钟,加入砂仁,再煎约6～8分钟,过滤,弃去滤渣。合并两次药液,浓缩直到需要的药液量(含生药1g/ml),分装,消毒,4℃储存备用。
3.治疗方法
生理盐水治疗对照组、藓化饮治疗组分别以生理盐水和中药藓化饮汤剂灌胃治疗,1.6ml/只,每天一次,持续八周,处死全部动物并取材,固定,镜检。
4.免疫组化法检测P53及COX-2在各组中的表达
取各组蜡块,常规石蜡切片,脱蜡至水,按照S-P试剂盒操作说明进行操作,镜下观察空白对照组,癌前病变对照组,生理盐水治疗对照组及藓化饮治疗组中的P53和COX-2的表达情况,并分析对比各组之间的表达情况是否存在差别,以评价中药藓化饮对口腔黏膜癌前病变的逆转作用。
5.流式细胞术
石蜡包埋组织在切片机上连续切取40-50μm厚的组织片5-8片,放入试管中;二甲苯脱蜡,梯度酒精水化;生理盐水冲洗,120目铜网轻搓组织,收集单细胞悬液;500目尼龙网过滤,离心,1000rpm,5分钟;加入一抗工作液,室温下静置40分钟;加入二抗工作液,室温下静置1小时;机检,读取结果。
6.统计学方法
所有采用SPSS 13.0统计软件,等级资料应用秩和检验方法分析,计量资料以均数±标准差(±S)表示,应用单因素方差分析对P53及COX-2检测结果进行统计学分析,α=0.05,P<0.05具有统计学意义。
结果:
1.在癌前病变对照组、生理盐水治疗对照组及藓化饮治疗组50例标本中,随机抽取10例标本在光镜下观察,均发现上皮分层不规则,基底细胞极性丧失,滴状钉突,细胞核多形性,核质比例增加,细胞核增大,不典型分裂像,核仁增大、数量增加及核深染等不典型增生表现,可说明癌前病变模型制备成功。
2.免疫组化
2.1 P53在空白对照组中阴性表达;在癌前病变对照组及生理盐水治疗对照组中均为强阳性表达;在藓化饮治疗组中为弱阳性表达。各组间差异有统计学意义,癌前病变对照组和生理盐水治疗对照组间差异无统计学意义。
2.2 COX-2在空白对照组中阴性表达;在癌前病变对照组及生理盐水治疗对照组中均为强阳性表达;在藓化饮治疗组中为弱阳性表达。各组间差异有统计学意义,癌前病变对照组和生理盐水治疗对照组间差异无统计学意义。
3.流式细胞术
3.1 P53在各组中均有表达,空白对照组和藓化饮治疗组均为弱阳性表达,组间差异无统计学意义;癌前病变对照组及生理盐水治疗对照组中均为强阳性表达,组间差异无统计学意义;藓化饮治疗组与癌前病变对照组相比,组间差异有统计学意义。
3.2 COX-2在各组中均有表达,空白对照组和藓化饮治疗组均为弱阳性表达,组间差异无统计学意义;癌前病变对照组及生理盐水治疗对照组中均为强阳性表达,组间差异无统计学意义;藓化饮治疗组与癌前病变对照组相比,组间差异有统计学意义。
4.相关性研究表明:P53与COX-2在口腔癌前病变的发生发展过程中,存在明显正相关性。
结论:
1.应用诱癌剂DMBA,在一定的时间和频次下,可有效诱导金黄地鼠颊黏膜癌前病变。
2.P53与COX-2在口腔黏膜癌前病变的发生和发展中有着重要的意义,P53与COX-2在口腔癌前病变的发生发展过程中,存在明显正相关性。
3.中药藓化饮对口腔癌前病变具有明显治疗效果。
Objective: The lesion precancerous of oral mucosa is a kind of precancerous state,showing off atypical hyperplasia of epithelium of oral mucosa in different extent,but it’s not be the cancer.The new categorization of WHO give the definition of lesion precancerous as that:Precursor lesions are defined as altered epithelium with an increased likelihood for progression to squamous cell carcinoma(SCC).The altered epithelium shows a variety of cytological and archi-tectural changes that have traditionally been grouped under the term dysplasia. Currently accepted precancerous lesions and precancerous conditions, includingleukoplakia, erythema, lichen planus and oral submucous fibrosis, etc. Without timely treatment of precancerous lesions and left unchecked, will ultimately develop into cancer, seriously affect the treatment and prognosis, but also to patients a tremendous impact on physical and psychological. How to treat oral precancerous lesions, blocking its development to oral cancer, has become a hotspot of current research. At present for the oral treatment of precancerous lesions of large, mainly Western oral, systemic and local use of steroids and immunosuppressive agents, early surgical excision and traditional Chinese medicine treatment. But the efficacy of Western medicine instability,and hormones and immune inhibitors, although the early onset of fast but its role is not sustainable and side effects are large; the most previous tradition of Chinese medicine connect with application of local hormones,so there’s little news about Chinese medicine treatment of precancerous lesions onlyand little study to prove the treatment effect of the Chinese medicine. Chinese medicine in the treatment of precancerous lesions of oral mucosa has obvious advantages for the characteristics of precancerous lesions of oral mucosa: Chinese medicine samples wen, effective solid; oral decoction or proprietary Chinese medicine preparation, taking the method is simple, patient compliance is good, easy to accept; with fewer side effects than western medicine and so on. In view of the above advantages, traditional Chinese medicine in the treatment of precancerous lesions of the increasing attention has been paid. In this study, drinking herbal Xianhuayin of experimental animals treated with oral precancerous lesion intervention, and by flow cytometry and immunohistochemistry to observe the before and after treatment P53 and COX-2 expression, analysis, and explore the Xianhuayin of drinking on the oral mucosa reversal of the role of precancerous lesions. For future clinical applications, experimental theory.
Methods:
1. Animal model of oral precancerous lesions and group preparation
Syrian golden hamster get (SPF grade) 55, 6 to 8 weeks of age, body length of about 10cm, weight 90 ~ 110g, both male and female, from Beijing Tong Lihua Experimental Animal Technology Victoria Limited, the production license number: SCXK (Beijing) 2007-0001. In accordance with the principle of completely randomized into four groups: untreat control group, 5, precancerous lesions control group, 10, and saline-treat control group, 10, Xianhuayin-treat group, 30 to drink.
The untreat control group, the normal daily feeding, untouched, until the end of the experiment, together with the other groups killed and drawn.
Precancerous lesion control group, saline-treat control group, Xianhuayin-treat group are painted by 0.5% DMBA at the cheek pouch mucosa three times a week for six weeks.
2. The brew of traditional Chinese medicine decoction of Xianhuayin
Traditional Chinese medicine in the first set of square by adding about 8 to 10 times as much water, soak 40 minutes to 1 hour, Wu Huo heat with 20 to 30 minutes to boil and maintain for about 10 minutes, and then about Huo Jian-Wen After 10 minutes, filtering, juice; then add the equivalent of six times the amount of the remaining residues in water, heated by Wu Huo 20 to 30 minutes to boil, with the text Huojian about 10 minutes, adding Amomum villosum, boil about 6 ~ 8 minutes, filtering, abandoned to the residues. Merging the two liquid, concentrated liquid until the required amount (including crude drug 1g/ml), packaging and sterilization, 4℃stored until required.
3. Xianhuayin of traditional Chinese medicine treatment of drinking and saline gavage
Saline-treat control group and Xianhuayin-treat group were treated with saline and drink decoction of Chinese medicine Xianhuayin gavage treatment, 1.6ml / only once a day for eight weeks , killed all the animals and subjects, fixed, microscopic examination.
4. Immunohistochemical detection of P53 and COX-2 expression in all groups
Take wax blocks of each group, routine paraffin sections, dewaxing to water, in accordance with S-P kit instructions to operate, microscopic observation of the untreat control group, precancerous lesions control group,saline-treat control group and Xianhuayin-treat group, in the P53 and COX-2 expression, and analysis and comparison between the expression of each group whether there are differences, to evaluate the traditional Chinese medicine Xianhuayin precancerous lesions of oral mucosa reverse effect.
5. Flow cytometry
Paraffin-embedded tissues on a continuous cut in the slicing machine to take the organization of 40-50μm thick film 5-8 tablets, placed in a test tube; xylene dewaxing, gradient alcohol hydration; saline wash, 120 mesh copper mesh Qingcuo organizations to collect a single cell suspension; 500 mesh nylon mesh filtration, centrifugation, 1000rpm, 5 minutes; to join an anti-working solution at room temperature, put it aside for 40 minutes; to join two anti-working fluid, put it aside for 1 hour at room temperature; machine inspection, read the results.
6. Statistical Methods
Using SPSS 13.0 statistical software, test results are the mean±standard deviation (±S) said that the application of analytical methods rank sum test P53 and COX-2 test results were statistically analyzed,α= 0.05, P <0.05 statistically significant.
Results:
1. Probabilistic selected specimens of 10 cases observed in the optical microscope, are found in irregular epithelial stratification, basal cell polarity loss, drop-shaped nail protrusion, pleomorphic nuclei, nuclear-cytoplasmic ratio increased, the nucleus increases, not like a typical split , nucleolus increases, increase in the number and nuclear deep-dyed so the performance of atypical hyperplasia, may indicate precancerous lesion models were successfully constructed.
2. Immunohistochemistry
2.1P53 in the untreat control group, negative expression; in the precancerous lesions control group and saline-treat control group were strongly positive expression; in the Xianhuayin-treat group for the weak positive expression. Differences between the groups was statistically significant, precancerous lesions control group and saline-treat control group, no significant difference.
2.2COX-2 in the untreat control group, negative expression; in the precancerous lesions control group and saline-treat control group were strongly positive expression; in the Xianhuayin-treat group for the weak positive expression. Differences between the groups was statistically significant, precancerous lesions control group and saline-treat control group, no significant difference.
3. FCM
3.1P53 were expressed in all groups, the untreat control group and the Xianhuayin-treat group were weak expression, no statistically significant difference between the groups; the precancerous lesions control group and saline-treat control group were strongly positive expression, no statistically significant difference between the groups; untreat control group and precancerous lesions compared with the control group, statistically significant difference between the groups.
3.2COX-2 were expressed in all groups, the untreat control group and the Xianhuayin-treat group were weak expression, no statistically significant difference between the groups; the precancerous lesions control group and saline-treat control group were strongly positive expression, no statistically significant difference between the groups; untreat control group and precancerous lesions compared with the control group, statistically significant difference between the groups.
4. Correlation studies have shown that: P53 and COX-2 in oral precancerous lesions of the occurrence and development process, there is significant correlation.
Conclusion:
1.Applications lure cancer agent DMBA, in a certain time and frequency, it can be effectively induced hamster buccal mucosa precancerous lesions.
2.P53 and COX-2 in oral precancerous lesions in the occurrence and development of great significance, P53 and COX-2 in oral precancerous lesions in the occurrence and development process, there is significant correlation.
3.Medicine Xianhuayin in the treatment of precancerous lesions of drink has a noticeable effect.
方法:
1.口腔黏膜癌前病变动物模型制备及分组
取叙利亚金黄地鼠(SPF级)55只,6～8周龄,体长约10cm,体重90～110g,雌雄兼备,由北京维通利华实验动物技术有限公司提供,生产许可证编号:SCXK(京)2007-0001。按照完全随机化原则分为四组:空白对照组5只,癌前病变对照组10只,生理盐水治疗对照组10只,藓化饮治疗组30只。
空白对照组每日正常饲养,不做处理,直至实验结束,与其余组一同处死并取材。
癌前病变对照组、生理盐水治疗对照组、藓化饮治疗组均以0.5%DMBA涂抹双侧颊黏膜,每周三次,持续六周。
2.中药藓化饮煎剂的制备
先向组方中药中加入约8～10倍的水,浸泡40分钟～1小时,用武火加热20～30分钟至煮沸,保持约10分钟,再以文火煎约10分钟后,过滤,取汁;再加入相当于剩余滤渣量6倍的水,用武火加热20～30分钟至煮沸,用文火煎约10分钟,加入砂仁,再煎约6～8分钟,过滤,弃去滤渣。合并两次药液,浓缩直到需要的药液量(含生药1g/ml),分装,消毒,4℃储存备用。
3.治疗方法
生理盐水治疗对照组、藓化饮治疗组分别以生理盐水和中药藓化饮汤剂灌胃治疗,1.6ml/只,每天一次,持续八周,处死全部动物并取材,固定,镜检。
4.免疫组化法检测P53及COX-2在各组中的表达
取各组蜡块,常规石蜡切片,脱蜡至水,按照S-P试剂盒操作说明进行操作,镜下观察空白对照组,癌前病变对照组,生理盐水治疗对照组及藓化饮治疗组中的P53和COX-2的表达情况,并分析对比各组之间的表达情况是否存在差别,以评价中药藓化饮对口腔黏膜癌前病变的逆转作用。
5.流式细胞术
石蜡包埋组织在切片机上连续切取40-50μm厚的组织片5-8片,放入试管中;二甲苯脱蜡,梯度酒精水化;生理盐水冲洗,120目铜网轻搓组织,收集单细胞悬液;500目尼龙网过滤,离心,1000rpm,5分钟;加入一抗工作液,室温下静置40分钟;加入二抗工作液,室温下静置1小时;机检,读取结果。
6.统计学方法
所有采用SPSS 13.0统计软件,等级资料应用秩和检验方法分析,计量资料以均数±标准差(±S)表示,应用单因素方差分析对P53及COX-2检测结果进行统计学分析,α=0.05,P<0.05具有统计学意义。
结果:
1.在癌前病变对照组、生理盐水治疗对照组及藓化饮治疗组50例标本中,随机抽取10例标本在光镜下观察,均发现上皮分层不规则,基底细胞极性丧失,滴状钉突,细胞核多形性,核质比例增加,细胞核增大,不典型分裂像,核仁增大、数量增加及核深染等不典型增生表现,可说明癌前病变模型制备成功。
2.免疫组化
2.1 P53在空白对照组中阴性表达;在癌前病变对照组及生理盐水治疗对照组中均为强阳性表达;在藓化饮治疗组中为弱阳性表达。各组间差异有统计学意义,癌前病变对照组和生理盐水治疗对照组间差异无统计学意义。
2.2 COX-2在空白对照组中阴性表达;在癌前病变对照组及生理盐水治疗对照组中均为强阳性表达;在藓化饮治疗组中为弱阳性表达。各组间差异有统计学意义,癌前病变对照组和生理盐水治疗对照组间差异无统计学意义。
3.流式细胞术
3.1 P53在各组中均有表达,空白对照组和藓化饮治疗组均为弱阳性表达,组间差异无统计学意义;癌前病变对照组及生理盐水治疗对照组中均为强阳性表达,组间差异无统计学意义;藓化饮治疗组与癌前病变对照组相比,组间差异有统计学意义。
3.2 COX-2在各组中均有表达,空白对照组和藓化饮治疗组均为弱阳性表达,组间差异无统计学意义;癌前病变对照组及生理盐水治疗对照组中均为强阳性表达,组间差异无统计学意义;藓化饮治疗组与癌前病变对照组相比,组间差异有统计学意义。
4.相关性研究表明:P53与COX-2在口腔癌前病变的发生发展过程中,存在明显正相关性。
结论:
1.应用诱癌剂DMBA,在一定的时间和频次下,可有效诱导金黄地鼠颊黏膜癌前病变。
2.P53与COX-2在口腔黏膜癌前病变的发生和发展中有着重要的意义,P53与COX-2在口腔癌前病变的发生发展过程中,存在明显正相关性。
3.中药藓化饮对口腔癌前病变具有明显治疗效果。
Objective: The lesion precancerous of oral mucosa is a kind of precancerous state,showing off atypical hyperplasia of epithelium of oral mucosa in different extent,but it’s not be the cancer.The new categorization of WHO give the definition of lesion precancerous as that:Precursor lesions are defined as altered epithelium with an increased likelihood for progression to squamous cell carcinoma(SCC).The altered epithelium shows a variety of cytological and archi-tectural changes that have traditionally been grouped under the term dysplasia. Currently accepted precancerous lesions and precancerous conditions, includingleukoplakia, erythema, lichen planus and oral submucous fibrosis, etc. Without timely treatment of precancerous lesions and left unchecked, will ultimately develop into cancer, seriously affect the treatment and prognosis, but also to patients a tremendous impact on physical and psychological. How to treat oral precancerous lesions, blocking its development to oral cancer, has become a hotspot of current research. At present for the oral treatment of precancerous lesions of large, mainly Western oral, systemic and local use of steroids and immunosuppressive agents, early surgical excision and traditional Chinese medicine treatment. But the efficacy of Western medicine instability,and hormones and immune inhibitors, although the early onset of fast but its role is not sustainable and side effects are large; the most previous tradition of Chinese medicine connect with application of local hormones,so there’s little news about Chinese medicine treatment of precancerous lesions onlyand little study to prove the treatment effect of the Chinese medicine. Chinese medicine in the treatment of precancerous lesions of oral mucosa has obvious advantages for the characteristics of precancerous lesions of oral mucosa: Chinese medicine samples wen, effective solid; oral decoction or proprietary Chinese medicine preparation, taking the method is simple, patient compliance is good, easy to accept; with fewer side effects than western medicine and so on. In view of the above advantages, traditional Chinese medicine in the treatment of precancerous lesions of the increasing attention has been paid. In this study, drinking herbal Xianhuayin of experimental animals treated with oral precancerous lesion intervention, and by flow cytometry and immunohistochemistry to observe the before and after treatment P53 and COX-2 expression, analysis, and explore the Xianhuayin of drinking on the oral mucosa reversal of the role of precancerous lesions. For future clinical applications, experimental theory.
Methods:
1. Animal model of oral precancerous lesions and group preparation
Syrian golden hamster get (SPF grade) 55, 6 to 8 weeks of age, body length of about 10cm, weight 90 ~ 110g, both male and female, from Beijing Tong Lihua Experimental Animal Technology Victoria Limited, the production license number: SCXK (Beijing) 2007-0001. In accordance with the principle of completely randomized into four groups: untreat control group, 5, precancerous lesions control group, 10, and saline-treat control group, 10, Xianhuayin-treat group, 30 to drink.
The untreat control group, the normal daily feeding, untouched, until the end of the experiment, together with the other groups killed and drawn.
Precancerous lesion control group, saline-treat control group, Xianhuayin-treat group are painted by 0.5% DMBA at the cheek pouch mucosa three times a week for six weeks.
2. The brew of traditional Chinese medicine decoction of Xianhuayin
Traditional Chinese medicine in the first set of square by adding about 8 to 10 times as much water, soak 40 minutes to 1 hour, Wu Huo heat with 20 to 30 minutes to boil and maintain for about 10 minutes, and then about Huo Jian-Wen After 10 minutes, filtering, juice; then add the equivalent of six times the amount of the remaining residues in water, heated by Wu Huo 20 to 30 minutes to boil, with the text Huojian about 10 minutes, adding Amomum villosum, boil about 6 ~ 8 minutes, filtering, abandoned to the residues. Merging the two liquid, concentrated liquid until the required amount (including crude drug 1g/ml), packaging and sterilization, 4℃stored until required.
3. Xianhuayin of traditional Chinese medicine treatment of drinking and saline gavage
Saline-treat control group and Xianhuayin-treat group were treated with saline and drink decoction of Chinese medicine Xianhuayin gavage treatment, 1.6ml / only once a day for eight weeks , killed all the animals and subjects, fixed, microscopic examination.
4. Immunohistochemical detection of P53 and COX-2 expression in all groups
Take wax blocks of each group, routine paraffin sections, dewaxing to water, in accordance with S-P kit instructions to operate, microscopic observation of the untreat control group, precancerous lesions control group,saline-treat control group and Xianhuayin-treat group, in the P53 and COX-2 expression, and analysis and comparison between the expression of each group whether there are differences, to evaluate the traditional Chinese medicine Xianhuayin precancerous lesions of oral mucosa reverse effect.
5. Flow cytometry
Paraffin-embedded tissues on a continuous cut in the slicing machine to take the organization of 40-50μm thick film 5-8 tablets, placed in a test tube; xylene dewaxing, gradient alcohol hydration; saline wash, 120 mesh copper mesh Qingcuo organizations to collect a single cell suspension; 500 mesh nylon mesh filtration, centrifugation, 1000rpm, 5 minutes; to join an anti-working solution at room temperature, put it aside for 40 minutes; to join two anti-working fluid, put it aside for 1 hour at room temperature; machine inspection, read the results.
6. Statistical Methods
Using SPSS 13.0 statistical software, test results are the mean±standard deviation (±S) said that the application of analytical methods rank sum test P53 and COX-2 test results were statistically analyzed,α= 0.05, P <0.05 statistically significant.
Results:
1. Probabilistic selected specimens of 10 cases observed in the optical microscope, are found in irregular epithelial stratification, basal cell polarity loss, drop-shaped nail protrusion, pleomorphic nuclei, nuclear-cytoplasmic ratio increased, the nucleus increases, not like a typical split , nucleolus increases, increase in the number and nuclear deep-dyed so the performance of atypical hyperplasia, may indicate precancerous lesion models were successfully constructed.
2. Immunohistochemistry
2.1P53 in the untreat control group, negative expression; in the precancerous lesions control group and saline-treat control group were strongly positive expression; in the Xianhuayin-treat group for the weak positive expression. Differences between the groups was statistically significant, precancerous lesions control group and saline-treat control group, no significant difference.
2.2COX-2 in the untreat control group, negative expression; in the precancerous lesions control group and saline-treat control group were strongly positive expression; in the Xianhuayin-treat group for the weak positive expression. Differences between the groups was statistically significant, precancerous lesions control group and saline-treat control group, no significant difference.
3. FCM
3.1P53 were expressed in all groups, the untreat control group and the Xianhuayin-treat group were weak expression, no statistically significant difference between the groups; the precancerous lesions control group and saline-treat control group were strongly positive expression, no statistically significant difference between the groups; untreat control group and precancerous lesions compared with the control group, statistically significant difference between the groups.
3.2COX-2 were expressed in all groups, the untreat control group and the Xianhuayin-treat group were weak expression, no statistically significant difference between the groups; the precancerous lesions control group and saline-treat control group were strongly positive expression, no statistically significant difference between the groups; untreat control group and precancerous lesions compared with the control group, statistically significant difference between the groups.
4. Correlation studies have shown that: P53 and COX-2 in oral precancerous lesions of the occurrence and development process, there is significant correlation.
Conclusion:
1.Applications lure cancer agent DMBA, in a certain time and frequency, it can be effectively induced hamster buccal mucosa precancerous lesions.
2.P53 and COX-2 in oral precancerous lesions in the occurrence and development of great significance, P53 and COX-2 in oral precancerous lesions in the occurrence and development process, there is significant correlation.
3.Medicine Xianhuayin in the treatment of precancerous lesions of drink has a noticeable effect.
引文
1 Pindborg JJ, Reichart P, Smith CJ, et al. World Health Organization:histological typing of cancer and precancer of the oral mucosa[M]. Berlin: Springer-Verlag, 1997: 25-26
2 Moschel RC, Baird WM, Dipple A. Metabolic activation of the carcinogen
7, 12-dimethylben(a) anthracene for DNA binding[J]. Biochem Biophys Res Commun, 1977, 76: 1092-1098
3魏玺,黄焰. DMBA诱导大鼠乳腺癌模型及乳腺癌化学预防实验研究进展.现代肿瘤医学[J].2008, 16 (3) : 454-456
4李宁,陈晓欣,韩驰,等. DMBA诱发的地鼠口腔癌发病机制研究[J].癌变·畸变·突变. 2003, 15 (1) : 21-24
5 Mithani SK, Mydlarz WK, Grumbine FL, et al. Molecular genetics of premalignant oral lesions[J]. Oral Dis, 2007, 13 (2) : 126-133
6陈谦明,江潞,曾新.口腔黏膜癌变相关基因研究[J].中华口腔医学研究杂志, 2008, 2 (3) : 201-210
7陈屏.肿瘤相关基因研究进展[J].肿瘤基础与临床杂志, 2008, 21 (1) : 90-92
8江锦岗,严振球.口腔癌前病变标志物的研究现状.临床军医杂志, 2006, 34 (2) : 224-227
9何迎春,卢芳国,田道法,等. TgN(P53mt-LMP1)/HT小鼠鼻腔和鼻咽黏膜上皮细胞增殖与凋亡相关基因表达的关系.癌变,畸变,突变. 2008, 20 (5) : 363-366
10 Trigiante G, Lu X. ASPP [corrected] and cancer[J]. Nat Rev Cancer, 2006, 6 (3) : 217-226
11张飞功,刘瑞,陈丹磊,等. P53, P21waf1, Ki-67及PCNA在小肠间质瘤的表达及其意义.中国普通外科杂志, 2009, 18 (4) : 406-408
12 Lu X. P53 : a heavily dictated dictater of life and death[J]. Curr Opin Genet Dev, 2005, 15 (1) : 27-33
13赵今,周学东,马三成,等. P16, P53, Ki-67在口腔癌前病变表达及4年临床随访.临床口腔医学杂志, 2004, 20 (4) : 588-590
14 Tabor MP, Brakenhoff RF, Ruijter-Schippers HJ, et al. Multple head and neck tumors frequently originate from a single preneoplastic lesion. Am J Pathol, 2002, 161 (2) : 1051-1060
15张春敏,陈显久.口腔癌前病变癌变的标志性基因研究进展.中国药物与临床, 2008, 8, suppl : 28-31
16杨啊晶,王雪梅,李电东,等. P53及其信号通路在肿瘤耐药分子机制中的研究进展.中国新药杂志, 2007, 12 (1) : 7-11
17董亚东,吉庆春,罗宝刚,等. PTEN和P53在食管鳞癌中的表达及意义.河南外科学杂志, 2009, 15 (1) : 6-7
18刘伟,吴益怀,陈玉芬,等.胃黏膜癌前病变中P53和增殖细胞核抗原的表达及其临床意义.临床消化病杂志, 2009, 21 (5) : 289-290
19郑曦,马宏,吴钦穗. P53蛋白和ki-67蛋白在食管鳞癌、癌前病变的表达及其临床意义.福建医科大学学报, 2006, 40 (3) : 243-245
20杨红卫. COX-2调节的前列腺素在突触信号传递中的作用.生理科学进展. 2009, 40 (4) : 317-320
21陈望燕,邓刚,姚琦. COX-2基因及产物在人类喉癌组织中的表达临床耳鼻咽喉头颈外科杂志. 2008, 22 (12) : 352-355
22孙江燕. COX-2、HIF-1α在口腔鳞状细胞癌组织中的表达和意义.肿瘤基础与临床. 2008, 6, 21 (3) : 247-249
23王滔明,许建明,胡乃中,等. Survivin、COX-2和VEGF在胃癌中的表达及其与预后的意义[J].临床消化病杂志, 2007, 19 (1) : 35-39
24 Gunnarsson C, Jansson A, Holmlund B, et al. Expression of COX-2 and steroid converting enzymes in breast cancer[J]. Oncol Rep. 2006, 16 (2) : 219-224
25 Crowell PL, Schmidt CM, Yip-Schneider MT, et al. Cyclooxygen-ase-2 expression in hamster and human pancreatic neoplasia[J]. Neoplasia, 2006, 8 (6) : 437-445
26魏欣,张汉东. Survivin在口腔鳞癌组织中的表达及其与COX-2相关性研究[J]. 2005, 21 (11) : 669-671
27任国欣,郭伟,环氧酶-2在人舌鳞癌及癌前病变组织中的表达[J].口腔颌面外科杂志, 2004, 14 (2) : 100-103
28 Subbaramaiah K, Altorki N, Chung WJ, et al. Inhibition of cyclooxygenase-2 gene expression by P53. J Biol Chem, 1999, 274 (16) : 10911
29吴卉娟,刘少扬,江大琼,等.宫颈癌中COX-2与P53、E-cadherin蛋白表达的关系.临床与实验病理学杂志. 2004, 20 (5) : 568-571
30杨含金,王建飞,郑炜智,等. 53例食管癌COX-2、P53和PCNA表达及临床意义.肿瘤学杂志. 2009, 15 (2) : 134-135
31毛越苹,段朝晖,曾凡钦,等.表皮肿瘤中COX-2与P53的检测.中国皮肤性病学杂志. 2006, 20 (11) : 668-670
32喻朝霞,戴瑜珍,胡小安,等.环氧化酶-2与P53、C-erbB-2、E-cadherin在胆囊癌中的表达及意义.安庆医学. 2006, 27 (1) : 8-11
33严蕊琳,谢永红.胃癌组织中COX-2表达及其与临床病理特征、P53、PCNA的关系.浙江预防医学. 2006, 18 (12) : 13-18
34朱群强,翦新春,凌天牖,等.中药2号口服液治疗口腔扁平苔藓前后人白细胞分化抗原DR区的表达变化,口腔医学2005, 25 (3) : 137～139
35徐学良,张梅,周汝俊,等.中药治疗口腔扁平苔藓的临床观察.临床口腔医学杂志, 1999, 15 (2) : 109～110
36许彦枝,尚丽乔,刘铁军,等.藓化饮对口腔黏膜癌前病变的逆转作用.肿瘤防治研究, 2008, 35 (12) : 874-877
37廖福龙,黄深,李文,等.活血化瘀药物药性的血流变学研究.中西医结合杂志, 1986, 6 (2) : 103
38袁伯勇.蝉蜕中氨基酸及微量元素分析.中国中药杂志. 1999, 15 : 35-36
39董昆山,王秀琴,董一凡.现代临床中药学.北京:中国中医药出版社, 1998. 471-531
1 Barnes L, Eveson JW, Reichart P, et al. Head and neck tumors[M]. Lyon : IARC Press, 2005 : 177-181
2 Warnakulasuriya S, Johnson NW, van der Waal I. Nomenclature and classification of potentially malignant disorders of the oral mucosa[J]. J Oral Pathol Med, 2007, 36 (10) : 575-580
3 Pindborg JJ, Reichart P, Smith CJ, et al. World Health Organization : histological typing of cancer and precancer of the oral mucosa[M]. Berlin : Springer-Verlag, 1997 : 25-26
4谢立群,沈丽佳,蒋会勇,等.组织芯片分析CD44v6和PCNA在口腔鳞癌及癌前病变中的表达及意义.暨南大学学报(医学版), 2006, 27 (6) : 767-772
5高黎,王国芳,季旭东,等.口腔黏膜白斑及鳞癌组织中核因子-κB p65与PCNA的表达.郑州大学学报(医学版), 2009, 44 (3) : 582-584
6范宗民,陈屏,刘小莉,等.食管癌变过程中p53、BrdU、P21waf1和PCNA蛋白表达的变化.郑州大学学报(医学版), 2006, 41 (1) : 27-29
7何迎春,卢芳国,田道法,等. TgN(p53mt-LMP1)/HT小鼠鼻腔和鼻咽黏膜上皮细胞增殖与凋亡相关基因表达的关系.癌变,畸变,突变. 2008, 20 (5) : 363-366
8徐志彬,吴明利,王士杰,等. PCNA、Bcl-2在家兔颊黏膜癌前病变制备及其逆转后的表达. 2008第五届中圈肿瘤学术大会论文集: 517
9 Gerdes J, Schwab U, Lemke H. et a1. Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation.Int J Caner, 1983. 31 : 13-15
10李媛,陆伟,王苏豫,等. CD147和Ki-67在口腔黏膜白斑和鳞癌组织中的表达及意义.中华老年口腔医学杂志, 2009, 7 (3) : 129-131
11李超,周洋. Ki-67与肺癌的临床研究进展.重庆医学, 2008, 37 (6) :647-649
12 Jalava P, Kuopio T, Juntti-Patinen L, et al. Ki-67 immunohistochemistry: a valuable marker in prognostication but with a risk of misclassification: proliferation sub-groups formed based on Ki-67 immunoreactivity and standardized mitotic index[J]. Histopathology, 2006, 48: 674
13马三成,赵今, Paul Speight. P53、Ki-67、P16和cyclin D1在口腔癌前病变的表达及相关性研究.苏州大学学报, 2009, 29 (3) : 480-482
14赵尔扬,方明,施磊,等.增殖细胞核抗原和Ki67在口腔癌浸润前沿表达及意义.口腔医学研究. 2008, 24 (5) : 495-497
15 Chulada PC, Thompson MB, Mahler JF, et al. Geneticdisruption of Ptags-l, aswellasPtgs-2, reduce sintestinal tumorigenesisin Minmice[J]. Cancer Res, 2000, 60(17) : 4705-4708
16杨红卫. COX-2调节的前列腺素在突触信号传递中的作用.生理科学进展. 2009, 40 (4) : 317-320
17陈望燕,邓刚,姚琦,等. COX-2基因及产物在人类喉癌组织中的表达临床耳鼻咽喉头颈外科杂志. 2008, 22(12) : 352-355
18孙江燕. COX-2、HIF-1α在口腔鳞状细胞癌组织中的表达和意义.肿瘤基础与临床. 2008, 6, 21 (3) : 247-249
19王滔明,许建明,胡乃中,等. Survivin、COX-2和VEGF在胃癌中的表达及其与预后的意义[J].临床消化病杂志, 2007, 19 (1): 35-39
20 Gunnarsson C, Jansson A, Holmlund B, et al. Expression of COX-2 and steroid converting enzymes in breast cancer[J]. OncolRep, 2006, 16 (2) : 219-224
21 CrowellPL, SchmidtCM, Yip-SchneiderMT, et al. Cyclooxygen-ase-2 expression in hamster and human pancreatic neoplasia[J]. Neoplasia, 2006, 8 (6) : 437-445
22 Sudbo J, Ristimaki A, Sondresen J E, et al. Cyclooxy-genase-2 (COX2) expression in high-risk premalignant oral lesions[J]. Oral Oncol, 2003, 39 : 497-505
23吴洪儒,孙正,汤晓飞,等. COX-2在口腔白斑、口腔扁平苔藓及口腔鳞状细胞癌中的表达.首都医科大学学报, 2006, 27(3) : 283-286
24龙智.一氧化氮(NO)与前列腺癌[J].国外医学:泌尿系统分册, 2003,22 (3) : 234-236
25黄小梅,郑兰荣,顾倩,等.口腔鳞癌及癌前病变组织中iNOS基因的表达.癌变,畸变,突变, 2008, 20 (1) : 50-55
26方豪,张水龙,许励,等.一氧化氮与口腔黏膜癌前病变的相关性研究.医学研究杂志, 2007, 36 (4) : 36-38
27 AltieriDC. Survivin, versatilemodulation ofcelldivision and apoptosis in cancer[J]. Oncogene, 2003, 22 (53) : 8581-8584
28 Altieri DC. Survivin, cancer networks and pathway-directed drug discovery. Nat Rev Cancer, 2008, 8 (1) : 61-70
29苏优勒,武文元. Survivin和Fhit基因与肿瘤研究进展.内蒙古民族大学学报, 2009, 15 (5) : 167-130
30都昌胡,徐军.凋亡蛋白抑制剂家族研究进展[J].世界华人消化杂志, 2005, 13 (13) : 1581-1589
31毛兰萍,刘晓锦,刘沛,等. Survivin,P21在人口腔扁平苔藓及口腔鳞癌中的表达.第四军医大学学报,2009,30 (21) : 2441-2443
32蒋鸿超,李鸿钧,孙茂盛.靶向Survivin肿瘤治疗研究进展[J].医学综述, 2006, 12 (3) : 163-166
33张春敏,陈显久.口腔癌前病变癌变的标志性基因研究进展.中国药物与临床, 2008, 8, suppl : 28-31
34赫丽杰,刘云鹏,侯科佐,等.胃癌组织中Survivin、COX-2蛋白表达及其与肿瘤血管生成的关系.现代肿瘤医学, 2009, 17 (9) : 1722-1725
35李祥勇,林观平,周克元.抑癌基因PTEN与细胞凋亡关系的研究进展[J].广州医学院学报, 2005, 23 (6) : 676-678
36 Li J, Yen C, Liaw D, et al. PTEN, a putative protein tyrosin phosphatase gene mutated in human brain,breast, prostate cancer[J]. Sciense, 1997, 275:1943
37 Shen Y H, Zhang L, Urama B, et al. Human cytomegalovirus inhibits Akt-mediated eNOS activation through upregulating PTEN[J]. Cardiovasc Res, 2006, 69 (2) : 502
38 Park KK, Liu K, Hu Y,et al. Promoting axon regeneration in the adultCNS by modulation of the PTEN/mTOR pathway[J]. Science, 2008, 322 (5903) : 963
39韩宇,曹青山,常廷民,等.食管癌中抑癌基因PTEN的表达及临床意义[J].新乡医学院学报, 2007, 24 (2) : 109-112
40林海.口腔扁平苔藓临床疗效研究中的问题及探讨[J].中华口腔医学杂志, 2005, 40 (2) : 105
41钟来平,郑家伟,张陈平,等.口腔癌早期诊断的研究现状[J].中国口腔颌面外科杂志, 2007, 5 (4) : 243
42林杰成,李志雄,林文霖.抑癌基因PTEN与食管癌关系研究进展[J].现代肿瘤医学, 2008, 5 : 844-847
43 Ohta M, Inoue H, Cotticelli M G, et al.The FHIT gene,spanning the chromosome 3p14.2 fragile site and renal carcinoma associated (3;8) breakpoint, is abnormal in digestive tract cancers[J]. Cell, 1996, 84 (4) 587-597
44 Suzuki A, Hayashida M, Ito T, et al.Survivin initiates cell cycle entry by the competitive interaction with Cdk4/p16(INK4a)and Cdk2/cyclin E complex activation[J]. Oncogene, 2006, 19 (29) : 3225-3234
45 Chaudhuri, Ito T, Kawano H, et al. Survivin initiates procaspase3/p21 complex formation as a result of interaction with Cdk4 to resist Fas-mediated cell death[J]. Oncogene, 2006, 19 (10) : 1346-1353
46殷操,沈丽佳,谢思明,等.口腔癌及癌前病变中FHIT与cyclinD1/CDK4表达的相关性研究.第一军医大学学报, 2005, 25 (7) : 812-814
47殷操,沈丽佳,谢思明,等.口腔鳞癌和癌前病变中脆性组氨酸三联体基因的表达及意义[J].第一军医大学学报, 2005, 25 (5) : 584-586
2 Moschel RC, Baird WM, Dipple A. Metabolic activation of the carcinogen
7, 12-dimethylben(a) anthracene for DNA binding[J]. Biochem Biophys Res Commun, 1977, 76: 1092-1098
3魏玺,黄焰. DMBA诱导大鼠乳腺癌模型及乳腺癌化学预防实验研究进展.现代肿瘤医学[J].2008, 16 (3) : 454-456
4李宁,陈晓欣,韩驰,等. DMBA诱发的地鼠口腔癌发病机制研究[J].癌变·畸变·突变. 2003, 15 (1) : 21-24
5 Mithani SK, Mydlarz WK, Grumbine FL, et al. Molecular genetics of premalignant oral lesions[J]. Oral Dis, 2007, 13 (2) : 126-133
6陈谦明,江潞,曾新.口腔黏膜癌变相关基因研究[J].中华口腔医学研究杂志, 2008, 2 (3) : 201-210
7陈屏.肿瘤相关基因研究进展[J].肿瘤基础与临床杂志, 2008, 21 (1) : 90-92
8江锦岗,严振球.口腔癌前病变标志物的研究现状.临床军医杂志, 2006, 34 (2) : 224-227
9何迎春,卢芳国,田道法,等. TgN(P53mt-LMP1)/HT小鼠鼻腔和鼻咽黏膜上皮细胞增殖与凋亡相关基因表达的关系.癌变,畸变,突变. 2008, 20 (5) : 363-366
10 Trigiante G, Lu X. ASPP [corrected] and cancer[J]. Nat Rev Cancer, 2006, 6 (3) : 217-226
11张飞功,刘瑞,陈丹磊,等. P53, P21waf1, Ki-67及PCNA在小肠间质瘤的表达及其意义.中国普通外科杂志, 2009, 18 (4) : 406-408
12 Lu X. P53 : a heavily dictated dictater of life and death[J]. Curr Opin Genet Dev, 2005, 15 (1) : 27-33
13赵今,周学东,马三成,等. P16, P53, Ki-67在口腔癌前病变表达及4年临床随访.临床口腔医学杂志, 2004, 20 (4) : 588-590
14 Tabor MP, Brakenhoff RF, Ruijter-Schippers HJ, et al. Multple head and neck tumors frequently originate from a single preneoplastic lesion. Am J Pathol, 2002, 161 (2) : 1051-1060
15张春敏,陈显久.口腔癌前病变癌变的标志性基因研究进展.中国药物与临床, 2008, 8, suppl : 28-31
16杨啊晶,王雪梅,李电东,等. P53及其信号通路在肿瘤耐药分子机制中的研究进展.中国新药杂志, 2007, 12 (1) : 7-11
17董亚东,吉庆春,罗宝刚,等. PTEN和P53在食管鳞癌中的表达及意义.河南外科学杂志, 2009, 15 (1) : 6-7
18刘伟,吴益怀,陈玉芬,等.胃黏膜癌前病变中P53和增殖细胞核抗原的表达及其临床意义.临床消化病杂志, 2009, 21 (5) : 289-290
19郑曦,马宏,吴钦穗. P53蛋白和ki-67蛋白在食管鳞癌、癌前病变的表达及其临床意义.福建医科大学学报, 2006, 40 (3) : 243-245
20杨红卫. COX-2调节的前列腺素在突触信号传递中的作用.生理科学进展. 2009, 40 (4) : 317-320
21陈望燕,邓刚,姚琦. COX-2基因及产物在人类喉癌组织中的表达临床耳鼻咽喉头颈外科杂志. 2008, 22 (12) : 352-355
22孙江燕. COX-2、HIF-1α在口腔鳞状细胞癌组织中的表达和意义.肿瘤基础与临床. 2008, 6, 21 (3) : 247-249
23王滔明,许建明,胡乃中,等. Survivin、COX-2和VEGF在胃癌中的表达及其与预后的意义[J].临床消化病杂志, 2007, 19 (1) : 35-39
24 Gunnarsson C, Jansson A, Holmlund B, et al. Expression of COX-2 and steroid converting enzymes in breast cancer[J]. Oncol Rep. 2006, 16 (2) : 219-224
25 Crowell PL, Schmidt CM, Yip-Schneider MT, et al. Cyclooxygen-ase-2 expression in hamster and human pancreatic neoplasia[J]. Neoplasia, 2006, 8 (6) : 437-445
26魏欣,张汉东. Survivin在口腔鳞癌组织中的表达及其与COX-2相关性研究[J]. 2005, 21 (11) : 669-671
27任国欣,郭伟,环氧酶-2在人舌鳞癌及癌前病变组织中的表达[J].口腔颌面外科杂志, 2004, 14 (2) : 100-103
28 Subbaramaiah K, Altorki N, Chung WJ, et al. Inhibition of cyclooxygenase-2 gene expression by P53. J Biol Chem, 1999, 274 (16) : 10911
29吴卉娟,刘少扬,江大琼,等.宫颈癌中COX-2与P53、E-cadherin蛋白表达的关系.临床与实验病理学杂志. 2004, 20 (5) : 568-571
30杨含金,王建飞,郑炜智,等. 53例食管癌COX-2、P53和PCNA表达及临床意义.肿瘤学杂志. 2009, 15 (2) : 134-135
31毛越苹,段朝晖,曾凡钦,等.表皮肿瘤中COX-2与P53的检测.中国皮肤性病学杂志. 2006, 20 (11) : 668-670
32喻朝霞,戴瑜珍,胡小安,等.环氧化酶-2与P53、C-erbB-2、E-cadherin在胆囊癌中的表达及意义.安庆医学. 2006, 27 (1) : 8-11
33严蕊琳,谢永红.胃癌组织中COX-2表达及其与临床病理特征、P53、PCNA的关系.浙江预防医学. 2006, 18 (12) : 13-18
34朱群强,翦新春,凌天牖,等.中药2号口服液治疗口腔扁平苔藓前后人白细胞分化抗原DR区的表达变化,口腔医学2005, 25 (3) : 137～139
35徐学良,张梅,周汝俊,等.中药治疗口腔扁平苔藓的临床观察.临床口腔医学杂志, 1999, 15 (2) : 109～110
36许彦枝,尚丽乔,刘铁军,等.藓化饮对口腔黏膜癌前病变的逆转作用.肿瘤防治研究, 2008, 35 (12) : 874-877
37廖福龙,黄深,李文,等.活血化瘀药物药性的血流变学研究.中西医结合杂志, 1986, 6 (2) : 103
38袁伯勇.蝉蜕中氨基酸及微量元素分析.中国中药杂志. 1999, 15 : 35-36
39董昆山,王秀琴,董一凡.现代临床中药学.北京:中国中医药出版社, 1998. 471-531
1 Barnes L, Eveson JW, Reichart P, et al. Head and neck tumors[M]. Lyon : IARC Press, 2005 : 177-181
2 Warnakulasuriya S, Johnson NW, van der Waal I. Nomenclature and classification of potentially malignant disorders of the oral mucosa[J]. J Oral Pathol Med, 2007, 36 (10) : 575-580
3 Pindborg JJ, Reichart P, Smith CJ, et al. World Health Organization : histological typing of cancer and precancer of the oral mucosa[M]. Berlin : Springer-Verlag, 1997 : 25-26
4谢立群,沈丽佳,蒋会勇,等.组织芯片分析CD44v6和PCNA在口腔鳞癌及癌前病变中的表达及意义.暨南大学学报(医学版), 2006, 27 (6) : 767-772
5高黎,王国芳,季旭东,等.口腔黏膜白斑及鳞癌组织中核因子-κB p65与PCNA的表达.郑州大学学报(医学版), 2009, 44 (3) : 582-584
6范宗民,陈屏,刘小莉,等.食管癌变过程中p53、BrdU、P21waf1和PCNA蛋白表达的变化.郑州大学学报(医学版), 2006, 41 (1) : 27-29
7何迎春,卢芳国,田道法,等. TgN(p53mt-LMP1)/HT小鼠鼻腔和鼻咽黏膜上皮细胞增殖与凋亡相关基因表达的关系.癌变,畸变,突变. 2008, 20 (5) : 363-366
8徐志彬,吴明利,王士杰,等. PCNA、Bcl-2在家兔颊黏膜癌前病变制备及其逆转后的表达. 2008第五届中圈肿瘤学术大会论文集: 517
9 Gerdes J, Schwab U, Lemke H. et a1. Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation.Int J Caner, 1983. 31 : 13-15
10李媛,陆伟,王苏豫,等. CD147和Ki-67在口腔黏膜白斑和鳞癌组织中的表达及意义.中华老年口腔医学杂志, 2009, 7 (3) : 129-131
11李超,周洋. Ki-67与肺癌的临床研究进展.重庆医学, 2008, 37 (6) :647-649
12 Jalava P, Kuopio T, Juntti-Patinen L, et al. Ki-67 immunohistochemistry: a valuable marker in prognostication but with a risk of misclassification: proliferation sub-groups formed based on Ki-67 immunoreactivity and standardized mitotic index[J]. Histopathology, 2006, 48: 674
13马三成,赵今, Paul Speight. P53、Ki-67、P16和cyclin D1在口腔癌前病变的表达及相关性研究.苏州大学学报, 2009, 29 (3) : 480-482
14赵尔扬,方明,施磊,等.增殖细胞核抗原和Ki67在口腔癌浸润前沿表达及意义.口腔医学研究. 2008, 24 (5) : 495-497
15 Chulada PC, Thompson MB, Mahler JF, et al. Geneticdisruption of Ptags-l, aswellasPtgs-2, reduce sintestinal tumorigenesisin Minmice[J]. Cancer Res, 2000, 60(17) : 4705-4708
16杨红卫. COX-2调节的前列腺素在突触信号传递中的作用.生理科学进展. 2009, 40 (4) : 317-320
17陈望燕,邓刚,姚琦,等. COX-2基因及产物在人类喉癌组织中的表达临床耳鼻咽喉头颈外科杂志. 2008, 22(12) : 352-355
18孙江燕. COX-2、HIF-1α在口腔鳞状细胞癌组织中的表达和意义.肿瘤基础与临床. 2008, 6, 21 (3) : 247-249
19王滔明,许建明,胡乃中,等. Survivin、COX-2和VEGF在胃癌中的表达及其与预后的意义[J].临床消化病杂志, 2007, 19 (1): 35-39
20 Gunnarsson C, Jansson A, Holmlund B, et al. Expression of COX-2 and steroid converting enzymes in breast cancer[J]. OncolRep, 2006, 16 (2) : 219-224
21 CrowellPL, SchmidtCM, Yip-SchneiderMT, et al. Cyclooxygen-ase-2 expression in hamster and human pancreatic neoplasia[J]. Neoplasia, 2006, 8 (6) : 437-445
22 Sudbo J, Ristimaki A, Sondresen J E, et al. Cyclooxy-genase-2 (COX2) expression in high-risk premalignant oral lesions[J]. Oral Oncol, 2003, 39 : 497-505
23吴洪儒,孙正,汤晓飞,等. COX-2在口腔白斑、口腔扁平苔藓及口腔鳞状细胞癌中的表达.首都医科大学学报, 2006, 27(3) : 283-286
24龙智.一氧化氮(NO)与前列腺癌[J].国外医学:泌尿系统分册, 2003,22 (3) : 234-236
25黄小梅,郑兰荣,顾倩,等.口腔鳞癌及癌前病变组织中iNOS基因的表达.癌变,畸变,突变, 2008, 20 (1) : 50-55
26方豪,张水龙,许励,等.一氧化氮与口腔黏膜癌前病变的相关性研究.医学研究杂志, 2007, 36 (4) : 36-38
27 AltieriDC. Survivin, versatilemodulation ofcelldivision and apoptosis in cancer[J]. Oncogene, 2003, 22 (53) : 8581-8584
28 Altieri DC. Survivin, cancer networks and pathway-directed drug discovery. Nat Rev Cancer, 2008, 8 (1) : 61-70
29苏优勒,武文元. Survivin和Fhit基因与肿瘤研究进展.内蒙古民族大学学报, 2009, 15 (5) : 167-130
30都昌胡,徐军.凋亡蛋白抑制剂家族研究进展[J].世界华人消化杂志, 2005, 13 (13) : 1581-1589
31毛兰萍,刘晓锦,刘沛,等. Survivin,P21在人口腔扁平苔藓及口腔鳞癌中的表达.第四军医大学学报,2009,30 (21) : 2441-2443
32蒋鸿超,李鸿钧,孙茂盛.靶向Survivin肿瘤治疗研究进展[J].医学综述, 2006, 12 (3) : 163-166
33张春敏,陈显久.口腔癌前病变癌变的标志性基因研究进展.中国药物与临床, 2008, 8, suppl : 28-31
34赫丽杰,刘云鹏,侯科佐,等.胃癌组织中Survivin、COX-2蛋白表达及其与肿瘤血管生成的关系.现代肿瘤医学, 2009, 17 (9) : 1722-1725
35李祥勇,林观平,周克元.抑癌基因PTEN与细胞凋亡关系的研究进展[J].广州医学院学报, 2005, 23 (6) : 676-678
36 Li J, Yen C, Liaw D, et al. PTEN, a putative protein tyrosin phosphatase gene mutated in human brain,breast, prostate cancer[J]. Sciense, 1997, 275:1943
37 Shen Y H, Zhang L, Urama B, et al. Human cytomegalovirus inhibits Akt-mediated eNOS activation through upregulating PTEN[J]. Cardiovasc Res, 2006, 69 (2) : 502
38 Park KK, Liu K, Hu Y,et al. Promoting axon regeneration in the adultCNS by modulation of the PTEN/mTOR pathway[J]. Science, 2008, 322 (5903) : 963
39韩宇,曹青山,常廷民,等.食管癌中抑癌基因PTEN的表达及临床意义[J].新乡医学院学报, 2007, 24 (2) : 109-112
40林海.口腔扁平苔藓临床疗效研究中的问题及探讨[J].中华口腔医学杂志, 2005, 40 (2) : 105
41钟来平,郑家伟,张陈平,等.口腔癌早期诊断的研究现状[J].中国口腔颌面外科杂志, 2007, 5 (4) : 243
42林杰成,李志雄,林文霖.抑癌基因PTEN与食管癌关系研究进展[J].现代肿瘤医学, 2008, 5 : 844-847
43 Ohta M, Inoue H, Cotticelli M G, et al.The FHIT gene,spanning the chromosome 3p14.2 fragile site and renal carcinoma associated (3;8) breakpoint, is abnormal in digestive tract cancers[J]. Cell, 1996, 84 (4) 587-597
44 Suzuki A, Hayashida M, Ito T, et al.Survivin initiates cell cycle entry by the competitive interaction with Cdk4/p16(INK4a)and Cdk2/cyclin E complex activation[J]. Oncogene, 2006, 19 (29) : 3225-3234
45 Chaudhuri, Ito T, Kawano H, et al. Survivin initiates procaspase3/p21 complex formation as a result of interaction with Cdk4 to resist Fas-mediated cell death[J]. Oncogene, 2006, 19 (10) : 1346-1353
46殷操,沈丽佳,谢思明,等.口腔癌及癌前病变中FHIT与cyclinD1/CDK4表达的相关性研究.第一军医大学学报, 2005, 25 (7) : 812-814
47殷操,沈丽佳,谢思明,等.口腔鳞癌和癌前病变中脆性组氨酸三联体基因的表达及意义[J].第一军医大学学报, 2005, 25 (5) : 584-586