| INTRODUCTIONLarynx as hormone target organ, its association to sex hormone as well as its receptor is paid close attention . Epidemiologic survey indicates that the incidence of laryngeal carcinoma in male is evidently higher than that in female, the ratio of men to women is 7 - 10 to 1 ,but we observe clinically that the characteristic of cancer of larynx in the northeast area of China is that the incidence of laryngeal cancer in female is evidently higher than that in other region of our country as well as many countries, and we discover with more survey that the ratio of men to women in supraglottic carcinoma of larynx is 1.3 to 1 as well as the ratio of men to women in glottic carcinoma of larynx is 11.8 to 1. Why is the incidence of the female in glottic carcinoma of larynx high? Whether or not it is relevant to sex hormone as well as its acceptor,there are not careful studies at present, so we want to do some explorations through experimentations in this aspect.MATERIALS AND METHODSMaterials: There are 63 specimens of laryngeal carcinoma,20 specimens of normal tissue beside carcinoma and 20 specimens of laryngeal benign lesion which were obtained by surgery in Department of Otolaryngology of our hospital. METHODS: In situ hybridization , all of these specimens are taken out from -70℃ low -temperature refrigerator, embeded by OCT again, cut by 6μm -thick tissue slice and place on the microscope slide disposed by APES. Placed in ordinary temperature in a minute, specimens are put into 4% paraformaldehyde again, then dried after they are flushed by distilled water. After they are take out , specimens are flushed by distilled water, then placed in mixed liquor of dioxo-gen and pure ethanol, flushed by distilled water after they are disposed in ordinary temperature. Pepsin are droped in specimens, and specimens are placed and digest in 38XI baking oven, then are flushed by PBS in situ hybridization and distilled water. Specimens are retained in ordinary temperature. Prehybrid-ization solution is placed on the slice of specimens,and incubated at 38°C baking oven for 3 hours. Then hybridization solution is placed, incubated at 38°C baking oven overnight. Next day the slices of specimens are taken out and flushed by 2 x SSCXO. 5 x SSCXO. 2 x SSC one by one in 37 °C . The slices of specimen are placed in 381 baking oven after confining liquid is placed in them. The slices of specimen are placed biotinylation anti - mice digoxin on and incubated at 38°C baking oven, then they are flushed by PBS in situ hybridization. The slices of specimen are placed SABC on and incubated at 38°Cbaking oven, then they are flushed thrice by PBS in situ hybridization. The slices of specimen are placed biotinylation peroxydase on and incubated at 38°Cbaking oven for 20 minutes, then followed by four wash with PBS in situ hybridization. The slices of specimen are placed DAB on for coloration, and washed by distilled water, stained by hematoxylin , thoroughly washed by water, washed by hydrochloric — acid alcohol for a moment, then soaked in the water after thoroughly washed by water, dewatered by alcohol in gradient,made transparent and mounting by neutral gum. The slices of specimens are observed by microscope. Those who are dyed buffy are masculine. Negative control: hybridization solution does not include hybridization probe of ER or AR, the others are same. Experimental datas are handled by X test.RESOULTSThe expressions of AR in male and female glottic carcinoma of larynx respectively are 77. 8% and 66.7% ,the expressions of AR in male and female su-praglottic carcinoma of larynx respectively are 66.7% and 60% ,the expressions of ER in male and female glottic carcinoma of larynx respectively are 66. 7% and 73.3% ,the expressions of ER in male and female supraglottic carcinoma of larynx respectively are 66.7% and 73. 3% .there are no significant difference between them( P >0. 05) ; there are no significant difference between the expressions of AR and ER in male and female carcinoma of larynx (P >0.05) ;the expressions of AR and ER in 20 specimens of normal tissue and 20 specimens of laryngeal benign lesion are 0% , there are significant differences compared with carcinoma of larynx( P < 0.01).DISCUSSIONIn our experiment, we apply in situ hybridization, which is such an important means for study of cell biology and molecular biology nowadays that experiment results are accurate and reliable.At present, larynx is already considered to be an target organ of sex hormone. Because there is a notable difference between male and female about the morbility of laryngeal carcinoma, many scholars believe laryngeal carcinoma is sex - hormone - dependent neoplasm.Some scholars believe that ER as well as its acceptor can prevent the growth of laryngeal carcinoma. Yang Beibei etc. believed that the ability of laryngeal carcinoma tissue expressing ER was acquired after canceration . Kleemann D etc. believed that female falling ill was concerned with decrease of ER after the onset of the climacteric phase and in the phase of senium, so ER was protection to larynx. Dai Chunfu etc. believed that ER was related to laryngeal carcinoma.Some scholars believe that AR as well as its acceptor can promote the growth of laryngeal carcinoma. Xu Wanchun etc. discovered that male of old age testosterone contents in serum were obviously higher than that in normal control, and the level of testosterone obviously decreased after removal of tumor. Through experiments Li Tianying etc. discovered that AR mRNA expressing in laryngeal carcinoma tissue was evidently higher than that in normal tissue beside carcinoma. Robin etc. witnessed that testosterone could promote the growth of laryngealcarcinoma cell , so they believed that AR in laryngeal carcinoma tissue supernor-mally requesting AR would promote the growth of tumor. Zhu Jianguo etc. believed that laryngeal carcinoma tissue not only reserved the ability of AR but also superaormally requested AR which would promote the growth of tumor and make tumor cells multiply rapidly, and he believed that protein content of AR in carcinoma tissue could be an index judging the level of degeneration and differentiation in tumor.Through experiments ,we discover that the expressions of AR in male and female glottic carcinoma of larynx respectively are 77. 8% and 66.1% ,the expressions of AR in male and female supraglottic carcinoma of larynx respectively are 66.7% and 60% ,the expressions of ER in male and female glottic carcinoma of larynx respectively are 66.1% and 73. 3% ,the expressions of ER in male and female supraglottic carcinoma of larynx respectively are 66. 7% and 73. 3% , there are no significant difference between them;there are no significant difference between the expressions of ARand ER in male and female carcinoma of larynx; the expressions of AR and ER in 20 specimens of normal tissue and 20 specimens of laryngeal benign lesion are 0% , there are significant differences compared with carcinoma of larynx. Experiment results above make clear that first of all , sex hormone receptor expressing is relevant to laryngeal carcinoma; next, sex hormone receptor expressing in laryngeal carcinoma does not differ in sex; what is more, AR and ER expressing in laryngeal carcimoma is all enhancing, which illustrates AR and ER simultaneously play a part in laryngeal carcimoma.Nowadays our experiment results illustrates that the expressions of sex her-mone acceptor are concerned with the incidence of carcinoma of larynx, but the difference in expression of AR and ER do not explain the difference of sex in incidence of laryngeal carcinoma, it show the significant difference of sex in the incidence of carcinoma of larynx may be concerned with other factors, which still needs us further research and study.
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