桥粒芯糖蛋白1（DSG1）在口腔粘膜白斑和口腔鳞癌中的表达
摘要
口腔白斑(oral leukoplakia, OLK)是指“是口腔黏膜上以白色为主的损害,不具有其他任何可定义的损害特征;一部分口腔白斑可转化为癌”。口腔粘膜上皮异常增生是口腔粘膜癌前病变的一个最基本特征,是上皮由正常状态向恶性转化的过渡阶段。由OLK向口腔鳞癌(oral squamous cell carcinoma, OSCC)的发展过程是多阶段、多步骤的。大量的研究表明其转变是一个受多因素影响的复杂过程。近来研究发现桥粒在肿瘤中的表达减少,尤其是在恶性肿瘤中基本不表达,桥粒在组织的恶变及肿瘤的转移中发挥了一定作用。本研究通过免疫组化技术检测口腔正常粘膜、口腔白斑、口腔鳞癌及体外培养的上皮细胞中桥粒芯糖蛋白1(desmoglein1, DSG1)的表达,以研究桥粒的变化规律,探讨其在癌变过程中的意义。
目的:
探讨桥粒在口腔粘膜癌变过程中的表达及意义,并且探讨口腔白斑癌变机制。
方法:
1.口腔正常上皮细胞体外培养,调节培养液中钙粒子浓度,免疫组化S-P法检测桥粒芯糖蛋白1(DSG1)的表达。口腔白斑成纤维细胞的培养:组织块法培养白斑成纤维细胞,绘制生长曲线,MTT法比较正常成纤维细胞与白斑成纤维细胞的生物学活力,免疫组化法检测角蛋白、波形蛋白表达。
2.口腔舌癌上皮细胞(Tca—8113)与口腔正常成纤维细胞、口腔白斑成纤维细胞在相同条件下混和培养3天,S-P法检测桥粒芯糖蛋白1(DSG1)的表达。
3.通过免疫组化S-P法检测来自山东大学口腔医院病理科1995—2002石蜡标本,包括正常口腔粘膜10例,OLK35例(其中单纯增生17例,轻度异常增生9例,中度异常增生6例,重度异常增生3例),OSCC30例(其中Ⅰ级12
Oral leukoplakia (OLK) do not possess the characteristics of any other lesion that can be defined , with white lesion on the oral mucosa, and Some can be turned into the cancer. The unusual hyperplasia of the mucous membrane is a essential feature of the precancer ,which is the transform stage from normal conditiont to malignan cancer. It is much stages , many steps from OLK to OSCC. Recent datas show that the expression of desmosome reduces in tumour, even no expression in malignant tumour especially.Desmosome play a certain role in the tumour cancerate and transformation. So we adopted immunohistochemical technique to study the expression of DSG1 in oral normal mucous ,OLK mucousa , OSCC mucousa and oral epithelium cultured in vitro.ObjectivesTo study the expression of Gsg1 in oral leukoplakia, and discuss the mechanism of the transformation from oral leukoplakia to carcinoma.Material and methods1. The expression of DSG1 were examined with immmuhistochemical SP method in the oral epithelium cultured in vitro, through regulating the density of calcium particle in medium. The primary OLK fibroblast was obtained by tissue culture. Drawing growth curve and measuring biological vigor by MTT method between OLK fibroblast and normal fibroblast .And the expression of cytokeratin, vimentin were examined with immmuhistochemical method .2. Tca—8113 cocultured with NFs, OLK fibroblast in the same condition for three
days. The expression of DSGl were examined with immmuhistochemical SP method inTca-8113.3. The expression of DSGl were examined with immmuhistochemical SP method in Paraffin wax sample which were from Shandong University stomatologicalhospital from 1995— 2002,including 10 normal oral mucous,35 OLK mucous(17 simple hyperplasia , 9 mild dysplasiarespectively,6 moderate dysplasiarespectively,3 severe dysplasiarespectively)and 30 oral squamous cell carcmomas(OSCC 112, OSCCII 10, OSCC III 8). And a half quantitative analysis was carried with the result ,and dealed with statistics.4. The changes were observed in different stage of canceration in OLK by transmission electronmlcroscope.Results1. We obtained oral epithelium and OLK fibroblast cultured in vitro. In the general ,the growth speed, proliferation and mitosis ability and vitality of the OLK fibroblast and NFs(p<0 .05). The oral OLK Fs showed negative staining for cytokeratin, and positive staining for vimentin .2. Result of EH: In normal oral mucosa, the positive rate of DSGl was 100%, and 94.1% in simple hyperplasia OLK, 27.8% in atypical hyperplasia OLK,13.3% in OSCC. There was positive result when the density of calcium particle in medium was 1.0 mM. There was negative result in Tca-8113,and there was positive result in Tca-8113 cocultured by OLK fibroblast.3. Result of OD: OD shrinked in order in in normal oral mucous, simple hyperplasia OLK, atypical hyperplasia OLK, OSCC. There were significant difference between each group(p<0.05).In OSCC, There were significant difference between each group(p<0.05).4. Result of transmission electronmlcroscope: In normal oral mucous there were plenty of desmosome with plenty of keratin intermediate filaments joined ,and nucleus was not destroyed. In canceration normal structure of desmosome were
destroyed, and keratinintermediate filaments were curled like balls, and nucleus was destroyed. ConclusionThe study shown that expression of DSG1 reduced in OLK especially in dysplasiarespectively OLK, and expression of DSG1 reduced more than OLK in OSCC ,and there is on expression in OSCCIII. This suggested that it play a role in the stage of OLK transferming to OSCC. This provides a new way for treating OLK and interdicting OLK cancerization.
目的:
探讨桥粒在口腔粘膜癌变过程中的表达及意义,并且探讨口腔白斑癌变机制。
方法:
1.口腔正常上皮细胞体外培养,调节培养液中钙粒子浓度,免疫组化S-P法检测桥粒芯糖蛋白1(DSG1)的表达。口腔白斑成纤维细胞的培养:组织块法培养白斑成纤维细胞,绘制生长曲线,MTT法比较正常成纤维细胞与白斑成纤维细胞的生物学活力,免疫组化法检测角蛋白、波形蛋白表达。
2.口腔舌癌上皮细胞(Tca—8113)与口腔正常成纤维细胞、口腔白斑成纤维细胞在相同条件下混和培养3天,S-P法检测桥粒芯糖蛋白1(DSG1)的表达。
3.通过免疫组化S-P法检测来自山东大学口腔医院病理科1995—2002石蜡标本,包括正常口腔粘膜10例,OLK35例(其中单纯增生17例,轻度异常增生9例,中度异常增生6例,重度异常增生3例),OSCC30例(其中Ⅰ级12
Oral leukoplakia (OLK) do not possess the characteristics of any other lesion that can be defined , with white lesion on the oral mucosa, and Some can be turned into the cancer. The unusual hyperplasia of the mucous membrane is a essential feature of the precancer ,which is the transform stage from normal conditiont to malignan cancer. It is much stages , many steps from OLK to OSCC. Recent datas show that the expression of desmosome reduces in tumour, even no expression in malignant tumour especially.Desmosome play a certain role in the tumour cancerate and transformation. So we adopted immunohistochemical technique to study the expression of DSG1 in oral normal mucous ,OLK mucousa , OSCC mucousa and oral epithelium cultured in vitro.ObjectivesTo study the expression of Gsg1 in oral leukoplakia, and discuss the mechanism of the transformation from oral leukoplakia to carcinoma.Material and methods1. The expression of DSG1 were examined with immmuhistochemical SP method in the oral epithelium cultured in vitro, through regulating the density of calcium particle in medium. The primary OLK fibroblast was obtained by tissue culture. Drawing growth curve and measuring biological vigor by MTT method between OLK fibroblast and normal fibroblast .And the expression of cytokeratin, vimentin were examined with immmuhistochemical method .2. Tca—8113 cocultured with NFs, OLK fibroblast in the same condition for three
days. The expression of DSGl were examined with immmuhistochemical SP method inTca-8113.3. The expression of DSGl were examined with immmuhistochemical SP method in Paraffin wax sample which were from Shandong University stomatologicalhospital from 1995— 2002,including 10 normal oral mucous,35 OLK mucous(17 simple hyperplasia , 9 mild dysplasiarespectively,6 moderate dysplasiarespectively,3 severe dysplasiarespectively)and 30 oral squamous cell carcmomas(OSCC 112, OSCCII 10, OSCC III 8). And a half quantitative analysis was carried with the result ,and dealed with statistics.4. The changes were observed in different stage of canceration in OLK by transmission electronmlcroscope.Results1. We obtained oral epithelium and OLK fibroblast cultured in vitro. In the general ,the growth speed, proliferation and mitosis ability and vitality of the OLK fibroblast and NFs(p<0 .05). The oral OLK Fs showed negative staining for cytokeratin, and positive staining for vimentin .2. Result of EH: In normal oral mucosa, the positive rate of DSGl was 100%, and 94.1% in simple hyperplasia OLK, 27.8% in atypical hyperplasia OLK,13.3% in OSCC. There was positive result when the density of calcium particle in medium was 1.0 mM. There was negative result in Tca-8113,and there was positive result in Tca-8113 cocultured by OLK fibroblast.3. Result of OD: OD shrinked in order in in normal oral mucous, simple hyperplasia OLK, atypical hyperplasia OLK, OSCC. There were significant difference between each group(p<0.05).In OSCC, There were significant difference between each group(p<0.05).4. Result of transmission electronmlcroscope: In normal oral mucous there were plenty of desmosome with plenty of keratin intermediate filaments joined ,and nucleus was not destroyed. In canceration normal structure of desmosome were
destroyed, and keratinintermediate filaments were curled like balls, and nucleus was destroyed. ConclusionThe study shown that expression of DSG1 reduced in OLK especially in dysplasiarespectively OLK, and expression of DSG1 reduced more than OLK in OSCC ,and there is on expression in OSCCIII. This suggested that it play a role in the stage of OLK transferming to OSCC. This provides a new way for treating OLK and interdicting OLK cancerization.
引文
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2.周曾同.提高口腔黏膜癌前病变的诊疗质量[J].中华口腔医学杂志,2005:40(2):89-91
3. Cawson RA. leukoplakia and oral cancer[J]. Dental Abstract, 1970; 15(1):32-35
4. Weir JC, Davenport WD, Skinner RL. A diagnostic and epidemiologic survey of 15,783 oral lesions [J]. Am Dent Assoc. 1987;115(3):439-42
5. Matsumoto M,Natsugoe S, Nakashima S, et al. Biological evaluation of undifferentiated carcinoma of the esophagus[J]. Ann Surg Oncol. 2000 ;7(3):204-9
6. Chow V, Yuen AP, Lam KY, et al. A comparative study of the clinicopathological significance of E-cadherin and catenins (alpha, beta, gamma) expression in the surgical management of oral tongue carcinoma[J]. Cancer Res Clin Oncol. 2001; 127(1):59-63
7. Miracco C, De Santi MM, Lio R, et al. Quantitatively evaluated ultrastructural findings can add to the differential diagnosis between keratoacanthoma and well differentiated squamous cell carcinoma[J]. Subrnicrosc Cytol Pathol. 1992 Jul;24(3):315-21
8. Oda D, Savard CE, Eng L, et al. Reconstituted human oral and esophageal mucosa in cuture[J]. In Vitro Cell Dev Biol Anita. 1998,34:46-52
9. Chang SE, Fosler S, Beess D, et al. Cultured gingival epithelium[J]. Cranio-Max-Fac Surg.1991,19:21-26
10. Krunic AL, Garrod DR, Madani S, et al. Immunohistochemical staining for desmogleins 1 and 2 in keratinocytic neoplasms with squamous phenotype: actinic keratosis, keratoacanthoma and squamous cell carcinoma of the skin[J]. Br J Cancer. 1998 Apr;77(8): 1275-9
11. Margrath I, Litvak J. Cancer in developing countries: opprunity and challenge [J]. Natl Cancer Inst. 1993, 85:862-874
12.张学军.桥粒与皮肤病[J].中华皮肤科杂志,2001,34(2):152-153
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