【作者】 刘明娜；

【导师】 吴先光；

【作者基本信息】 汕头大学 ， 耳鼻咽喉科学， 2011， 硕士

【摘要】 目的通过对HPV、CyclinD1、p15、EGFR和Bax蛋白在鼻腔、鼻窦内翻性乳头状瘤、鼻鳞状细胞癌及炎性鼻息肉组织中定位表达的比较,分析它们在鼻腔﹑鼻窦内翻性乳头状瘤和鼻鳞状细胞癌发生过程中的作用及相关意义,从而指导临床治疗。材料方法收集汕头大学医学院病理教研室及第二附属医院病理科1999年-2009年收检的鼻腔、鼻窦肿瘤临床资料保存完整的手术切除或鼻内活检组织存档蜡块60例,其中11例鼻炎性息肉(INP)、34例鼻腔、鼻窦内翻性乳头状瘤(SNIP)、15例鼻鳞状细胞癌(SCC)。采用免疫组织化学EnVision两步法,检测60例组织标本中HPV、CyclinD1、p15、EGFR及Bax蛋白的表达情况。SPSS17.0统计软件分析结果,各组两两比较用χ~2检验及Fisher精确检验法,各蛋白间相关性用Pearson相关分析法,检验水准α=0.05,P<0.05为差异有统计学意义,P<0.01为差异有高度统计学意义。结果1.病理观察结果:34例SNIP中3例为假复层纤毛柱状上皮乳头状瘤,其余为鳞状上皮乳头状瘤;3例上皮有中、重度不典型增生。15例SCC中有2例中同时含有鳞癌和内翻性乳头状瘤两种肿瘤组织,其余13例为单纯鳞状细胞癌。2.免疫组织化学检测结果:⑴光镜下见HPV蛋白阳性定位于细胞核,呈浅黄色或浅棕色;CyclinD1、p15蛋白阳性均定位于细胞核,呈棕黄或棕褐色;EGFR蛋白阳性定位于细胞膜,呈棕黄色;Bax蛋白阳性定位于细胞浆,呈浅黄色或深棕色。⑵①HPV蛋白在INP、SNIP、SCC组中阳性表达率为0%、38.2%、26.7%。SNIP与INP组间差异有统计学意义(χ~2=5.915,P=0.018<0.05),其余组间均无统计学意义(P>0.05)。②CyclinD1蛋白在INP、SNIP、SCC组中阳性表达率为18.18%、35.29%、66.67%。SCC与INP组间差异有统计学意义(χ~2=6.003,P=0.017<0.05),其余组间均无统计学意义(P>0.05)。③p15蛋白在INP、SNIP、SCC组中阳性表达率为90.91%、35.29%、26.67%。SNIP与INP组、SCC与INP组间比较,差异均有高度统计学意义(χ~2=10.288,P=0.001<0.01;χ~2=10.539,P=0.001<0.01),SCC与SNIP组间无统计学意义(P>0.05)。④EGFR蛋白在INP、SNIP、SCC组中阳性表达率为18.18%、79.41%、93.33%。SCC与INP组、SNIP与INP组间比较,差异均有高度统计学意义(χ~2=15.143,P=0.00016<0.01;χ~2=13.599,P=0.00048<0.01),SCC与SNIP组间无统计学意义(P>0.05)。⑤Bax蛋白在INP、SNIP、SCC组中阳性表达率为81.82%、70.59%、33.33%。SCC与SNP组、SCC与SNIP组间差异均有统计学意义(χ~2=6.003,P=0.017<0.05;χ~2=5.980,P=0.026<0.05),SNIP与INP组间无统计学意义(P>0.05)。⑥SNIP和SCC组中HPV、CyclinD1、p15、EGFR和Bax五种蛋白表达强度之间均无相关性(P>0.05)。结论1、HPV感染与鼻腔、鼻窦内翻性乳头状瘤的发生密切相关,进一步证实HPV感染是导致鼻腔、鼻窦内翻性乳头状瘤发生的重要病因学因素之一,可以作为鼻腔、鼻窦内翻性乳头状瘤发生的重要辅助指标;2、CyclinD1蛋白的过度表达与鼻鳞状细胞癌的发生密切相关,提示CyclinD1蛋白的过度表达可能在鼻鳞状细胞癌的发生过程中起着重要作用。CyclinD1蛋白的高表达可能对鼻腔、鼻窦内翻性乳头状瘤癌变有促进趋势,但这还需在扩大样本量进一步证实;3、p15蛋白低表达与鼻腔、鼻窦内翻性乳头状瘤以及鼻鳞状细胞癌发生和发展密切相关,提示p15蛋白的低表达可能在鼻腔、鼻窦内翻性乳头状瘤的发生及鼻鳞状细胞癌发生、发展复杂过程中有重要作用;4、EGFR蛋白的过度表达与鼻腔、鼻窦内翻性乳头状瘤以及鼻鳞状细胞癌发生和发展密切相关,提示EGFR蛋白的高表达可能在鼻腔、鼻窦内翻性乳头状瘤的发生及鼻鳞状细胞癌的发生过程中有重要作用。5、Bax蛋白显著低表达与鼻鳞状细胞癌的发生、发展过程密切相关,提示Bax蛋白低表达可能在鼻鳞状细胞癌的发生、发展过程中起着非常重要的负调控作用并且可能预示着鼻腔、鼻窦内翻性乳头状瘤有恶变倾向;6、HPV、CyclinD1、p15、EGFR和Bax五种蛋白在鼻腔、鼻窦内翻性乳头状瘤及鼻鳞状细胞癌中的表达无相关性,这有待于进一步研究;更多还原

【Abstract】 ObjectiveTo investigate the expression of HPV、CyclinD1、p15、EGFR and Bax proteins in thesinonasal inverted papilloma、nasal squamous cell carcinoma and inflammatory nasal polyptissues,study the roles and relationships in the occurrence process of the sinonasal invertedpapilloma and nasal squamous cell carcinoma.Materials and MethodsParaffin blocks of surgical specimen of the tumors of nasal cavity and sinus and intranasalbiopsy, including 11 cases of inflammatory nasal polyp(INP),34 of the sinonasal invertedpapilloma (SNIP),15 of nasal squamous cell carcinoma(SCC) were selected from Shantouuniversity medical college pathology staff room and Dept.of pathology of second affiliatedhospital from 1999 to 2009. Expression of HPV、CyclinD1、p15、EGFR and Bax proteins weredetected by immunohistochemistry Envision two-steps method in 60 tissue samples. Statisticalanalysis was performed using SPSS17.0. Chi-square test and Fisher’s exact test were used toobtain statistically significant differences between two groups. Pearson correlation coefficientswere calculated to evaluate the relationships among proteins. The correctedα=0.05. A value ofP<0.05 was considered statistically significant. A value of P<0.01 was considered highlystatiscally significant.Results1. Pathological observations：3 cases of 34 cases SNIP were pseudostratified ciliated columnar epithelium papilloma, therest were squamous papilloma; the epithelials of 3 cases had moderate and severe cervicalsquamous cell dysplasia. 1 case of 15 cases SCC was with a little inverted papillomaorganization, the rest 14 cases were simple squamous cell carcinoma.2.Immunohistochemical tests： ⑴Under the light microscope, the positive area of HPV protein was located in the nuclears,showing buff or shallow brown; the positive areas of CyclinD1 and p15 proteins were locatedin the nuclears, showing brown-yellow or brown; the positive areas of EGFR protein waslocated in the membranes, showing brown-yellow; positive areas of Bax protein was locatedin the cytoplasm showing light yellow or dark brown.⑵①Positive expression rates of HPV protein were respectively 0%、38.24%、26.67% in theINP、SNIP、SCC. The difference between SNIP and INP was statistically significant(χ2=5.915，P=0.018<0.05).The rest were no statistically significant among groups(P>0.05).②Positive expression rates of CyclinD1 protein were respectively 18.18%、35.29%、66.67% inthe INP、SNIP、SCC. The difference between SCC and INP was statistically significant(χ2=6.003，P=0.017<0.05).The rest were no statistically significant among groups(P>0.05).③Positive expression rates of p15 protein were respectively 90.91%、35.29%、26.67% in theINP、SNIP、SCC. The differences between SNIP and INP and between SCC and INP werehighly statistically significant(χ~2=10.288，P=0.001<0.01；χ~2=10.539，P=0.001<0.01).Thedifference between SCC and SNIP was no statistically significant (P>0.05).④Positive expression rates of EGFR protein were respectively 18.18%、79.41%、93.33% in theINP、SNIP、SCC. Significant differences were obtained between SCC and INP (χ~2=15.143，P=0.00016<0.01) and between SNIP and INP (χ2=13.599，P=0.00048<0.01).Thedifference between SCC and SNIP was no statistically significant (P>0.05).⑤Positive expression rates of Bax protein were respectively 81.82%、70.59%、33.33% in theINP、SNIP、SCC. The differences between SCC and INP and between SCC and SNIP werestatistically significan（tχ~2=6.003，P=0.017<0.05；χ~2=5.980，P=0.026<0.05）。The differencebetween SNIP and INP was no statistically significant (P>0.05).⑥No significant difference was found among the expression of HPV、CyclinD1、p15、Bax、EGFR five kinds of proteins in the SNIP and SCC（P>0.05）.Conclusions1. There is relationship between HPV infection and the occurrence of SNIP, to further prove that HPV infection is one of important etiological factors in the occurrence of SNIP and HPVis an important index in the occurrence of SNIP.2. There is relationship between over-expressed of CyclinD1 protein and SCC; it suggests thatover-expressed of CyclinD1 protein may play an important role in the process of theoccurrence of SCC. High expression of CyclinD1 protein may has the trend to promote thecancerous of SNIP, but it still needs to expand in sample size to further confirmation;3. There are relationships between low expression of p15 protein and SNIP and SCC; itsuggests that low expression of p15 protein may play important roles in the process of theoccurrence of SNIP and SCC;4. There are relationships between excessive expression of EGFR protein and SNIP and SCC; itsuggests that excessive expression of EGFR protein may play important roles in the processof the occurrence of SNIP and SCC;5. There is relationship between significantly low expression of Bax protein and SCC; itsuggests that low expression of Bax protein may have a very importantly negative regulatingrole in the canceration of SCC and indicate that SNIP has the trend of canceration;6. The expressions of HPV, CyclinD1, p15, Bax and EGFR protein have no correlations in theSNIP and SCC, and this needs to be further study;更多还原