胸部肿瘤患者肋间肌碳酸酐酶Ⅲ及其mRNA表达的研究

The study of the expression of carbonic anhydrase Ⅲ and its m RNA in the intercostals muscles of chest tumor patients

背景与目的:有研究提示肿瘤可以上调其原发灶及转移灶中的碳酸酐酶(carbonic anhydrase,CA)含量,但至目前为止尚不知肿瘤组织是否会影响其他非转移组织中CA的含量。本研究通过测定CAⅢ及其m RNA在胸部良性肿瘤、肺腺癌患者肋间肌中的表达,探讨CAⅢ在这些疾病状态下的表达调控情况。方法:对昆明医科大学第一附属医院胸外科行开胸肿瘤切除术的38例住院患者取肋间肌组织活检,根据术后肿瘤组织病理结果提示纵隔良性肿瘤10例、肺腺癌12例。肺腺癌组患者根据肿瘤转移情况分为淋巴结转移组(6例)和无淋巴结转移组(6例),根据分化程度分为低分化组(3例)、中分化组(6例)及高分化组(3例),此外,16例单纯胸外伤作为对照组。然后,利用ELISA法定量测定患者肋间肌CAⅢ的浓度,利用Real-time PCR法检测患者肋间肌的CAⅢm RNA表达水平,分析比较各组患者肋间肌中CAⅢ的浓度定量及其m RNA表达水平差异,探讨CAⅢ在肺腺癌等疾病状况下的表达调控情况。结果:CAⅢ的浓度定量结果方面,与其他2组比较,肺腺癌组患者肋间肌CAⅢ的浓度显著升高(P<0.01),且淋巴结转移组高于无淋巴结转移组(P<0.05),CAⅢ浓度在低、中、高分化肺腺癌各组患者肋间肌间的表达差异无统计学意义(P均>0.05)。在CAⅢm RNA相对表达量方面(2-△△Ct),与其他2组比较,肺腺癌组患者肋间肌CAⅢm RNA表达水平显著升高(P<0.01),且淋巴结转移组高于无淋巴结转移组高(P<0.01),但CAⅢm RNA水平在低、中、高分化肺腺癌各组患者肋间肌间差异无统计学意义(P均>0.05)。患者CAⅢ的浓度定量及其m RNA表达水平与患者的FEV1/Pred%(第一秒呼气容积占预计值百分比)呈正相关(P<0.05),与患者的年龄、体质量指数、FEV1/FVC%、Pa O2、Pa CO2无显著相关性(P均>0.05)。在各组患者,其肋间肌CAⅢm RNA表达水平与CAⅢ浓度定量呈现一致性表现,即m RNA表达水平升高组,其蛋白浓度定量也升高;相反,m RNA表达水平降低组,其蛋白浓度定量也降低。结论:肺腺癌组患者肋间肌的CAⅢ的浓度定量及其m RNA水平表达均较其他2组高。由于肺腺癌肋间肌CAⅢ的m RNA表达水平与CAⅢ的浓度定量呈现一致性表现,其对CAⅢ的调控可能主要在上游DNA转录水平得到实现,使得其肋间肌CAⅢm RNA转录水平及蛋白表达异常,进而影响其肋间肌功能状态。

Background and purpose： Many studies have showed that tumor tissues can up-regulate the con- tent of carbonic anhydrase （CA） at the primary lesion and metastasis, but, Until to now, we still did not know whether tumor tissues could affect the CA content at other non-metastasis tissues. In this study, we should detect the expression levels of CA]II and mRNA in the intercostals muscles of patients with chest tumor, to explore the regulation mechanism of CAm in patients with certain disease states. Methods： Intercostals muscle biopsy specimens were obtained from 38 in patients of the First Affiliated Hospital of Kunming Medical University. A total of 10 patients with benign mediastinal tumor, and 12 patients with lung adenocarcinoma were included according to the postoperative pathologic results. Lung adenoearcinoma group was divided into lymph node metastasis group （6 cases） and non-metastasis group （6 cases）, and divided into low differentiatiated carcinoma group （3 cases）, moderately differentiatiated carcinoma group （6 cases） and well differentiatiated carcinoma group （3 cases） and 16 cases purely chest trauma also were included as control group. The concentrations quantification of CAⅢ were measured by ELISA methods, the mRNA expression levels of CAⅢ were detected using RT-PCR methods. Results： The concentrations quantification of CAⅢ were remarkably raised in intercostals muscles of lung adenocarcinoma group compared to other two group （P〈0.05）, and the concentrations were tend to be higher in the group of lung adenocarcinoma with lymph node metastasis than the group of non-metas- tasis （P〈0.05）, but there were no significant difference of the concentrations of CAⅢ among each group of low dif- ferentiated adenocarcinoma, moderately differentiated adenocarcinoma and well differentiatiated adenocarcinoma （each P〉0.05）. The mRNA expression levels of CAⅢ were remarkably raised in intercostals muscles of lung adenocarci- noma group compared to control group （P〈0.01）, and the mRNA levels were tend to be higher in the group of lung adenocarcinoma with lymph node metastasis than the group of non-metastasis （P〈0.05）, but there were no significant difference of the mRNA levels of CA Ⅲ among each group of low differentiated adenocarcinoma, moderately differ- entiated adenocarcinoma and well differentiatiated adenocarcinoma （each P〉0.05）. The concentrations quantification and mRNA expression levels of CAⅢ in intercostals muscles were positively correlation with patient＇s FEVI/Pred% （P〈0.05）, but there were no significant correlation with patient＇s age, BMI, FEV1/FVC%, PaO2 and PaCO2 （P〉0.05）. This study showed a consistent performance between the mRNA expression levels and concentrations quantification of CAⅢ in intercostals muscles of each group. The higher mRNA expression levels of CA 11I in intercostals muscles of each group, the higher protein concentrations quantification; and the lower mRNA expression levels of CAⅢ, the lower protein concentration quantification. Conclusion： The concentrations quantification and mRNA expression levels of CAⅢ in intercostals muscles of lung adenocarcinoma group were higher than control group.This study showed a con- sistent performance between mRNA expression levels and concentrations quantification of CAⅢ in intercostals muscles of each group, it suggested that the regulation of CAⅢ in intercostals muscles of COPD patients might be achieved mainly in the upstream of DNA transcription level, which makes the transcription levels of mRNA and protein expres- sion of CA Ⅲ abnormal and further affects the state of intercostals muscles function.