Expression of Coxsackievirus and Adenovirus Receptor in Human Lung Cancer： Possible Clinical Significance

OBJECTIVE To explore the relationship between CAR and the development of human lung cancer, as well as to provide the basis for the clinical treatment of lung cancer using an adenovirus vector-based gene therapy. METHODS CAR expression was assessed immunohisto- chemically in tumoral, paraneoplastic and normal samples from 112 lung cancer patients. At the same time, the mRNA and protein expression of CAR in 32 cases were determined by RT-PCR and Western blot. The relationship between CAR expression and clinicopathologic parameters was statistically analyzed. RESULTS There was no expression of CAR in normal lung tissue but a little in paraneoplastic tissue. The positive rate was 43% in squamous cell carcinoma, and 70% in adenocarcinoma. Both were much significantly higher than that in paraneoplastic tissue. The CAR expression level in adenocarcinoma was higher than that in squamous cell cancer, mRNA expression by RT-PCR and protein expression by Western blot were consistent with immunohistochemistry results. CONCLUSION CAR is overexpressed in human lung cancer, especially in adenocarcinoma. This data offer the reliable basis for adenovirus-mediated gene therapy of lung cancer; more important, CAR may take part in the formation or development of lung cancer; this may be exploitable for the development of antibody-directed therapy in human lung cancer.

Expression of coxsackie and adenovirus receptor in small cell lung cancer

Objective: We explored the expression of coxsackie and adenovirus receptor (CAR) in small cell lung cancer (SCLC) tissue. Methods: CAR expression in 31 SCLC was assessed in formaldehyde-fixed, paraffin-embedded tissue according to the EnVision immunohistochemistry procedure, while 3 samples of surgical specimens of non-malignant lung disease were taken as the negative control. Results: We observed that the expression of CAR was detectable positive in all the 31 cases from the small cell lung cancer tissue, in contrasting that non-malignant lung tissue control. Conclusion: The high expression of CAR appeared in SCLC tissue indicates that it play an important role in of adenovirus vector-based gene therapy in SCLC.

Expression of Coxsackie and Adenovirurus Receptor and its Significance in Human Lung Cancer

OBJECTIVE To study the relationship between the coxsackie and adenovirus receptor （CAR） and the development of human lung cancer. To optimize adenovirus vector-based gene therapy.METHODS The expression of CAR in 112 cases of lung cancer was examined using immunohistochemistry. At the same time, the relationship between CAR expression and clinicopathologic characteristics was analyzed,RESULTS ：lhere is a little expression of CAR in normal lung tissue. Compared with paraneoplastic epithelial tissue of the lung, the expression of CAR is generally up-regulated in tumor tissues showing a significant dif- ference （P〈0.01）. The positive rate of CAR expression in squamous cell carcinoma was 43.1%, and in adenocarcinoma 70.2%, with the difference between the two rates being statistically significant （P〈0.01）. Compared to the paraneoplastic tissues, the difference in CAR positive expression was 35.4% for squamous cell carcinoma and 38.3% for adenocarcinoma. But the difference in different stages of squamous cell carcinoma had no statistical significance （P〉0.05）. However, the expression of CAR was at a high level in the bronchioalveolar carcinomas as 80.4% were CAR positive. This research showed that there was a specially high expression of CAR in adenocarcinomas.CONCLUSION CAR is expressed in human lungs at a low level and up-regulated in the tumor tissues, suggesting that there is a relationship between adenocarcinoma and CAR. This research provides a basis for planning a regimen of gene therapy using an adenovirus vector,

肿瘤细胞CAR表达水平与5型腺病毒转导效率的关系

目的：通过体外、体内实验探讨肿瘤细胞柯萨奇腺病毒受体（coxsackie adenovirus receptor，CAR）表达水平与腺病毒转导效率的关系，为腺病毒相关的生物制剂在临床个体化应用提供实验依据。方法：将载有绿色荧光蛋白的Ad5型腺病毒（Ad．GFP）以100MOI、200MOI分别感染人食管癌细胞系KYSE510、KYSE150、EC9706、人宫颈癌细胞系HeLa、人卵巢癌细胞系SKOV3、人肝癌细胞系HepG2和人肺癌细胞系A549，感染后48h通过流式细胞术检测Ad—GFP对不同细胞系的转导效率。采用Western blotting方法检测这些细胞中CAR的表达水平。将HeLa、A549、SKOV3、EC9706细胞分别接种于裸鼠腋下，接种细胞数分别为3×10^6、3×10^6、3×10^6、2×10^6，建立裸鼠移植瘤模型。待肿瘤长径达5～7mm时，在裸鼠移植瘤内注射Ad—GFP，每次1×10^9PFU，间隔48h注射第2次。第2次注射后48h处死裸鼠，剖取瘤组织，荧光显微镜观察冰冻切片中GFP的表达情况以判定腺病毒在瘤体内的转导效率，同时用免疫组化法检测瘤组织内的CAR表达水平。结果：200MOIAd—GFP感染A549、HeLa、HepG2、KYSE150细胞48h后分别有92．67％、89．31％、84．98％、74．59％的细胞表达GFP；而SKOV3、KYSE510、EC9706细胞中腺病毒的转导效率明显降低，GFP阳性率分别为30．06％、27．40％、18．93％；各种细胞的CAR蛋白表达水平与腺病毒的转导效率呈正相关。注射Ad—GFP的裸鼠移植瘤组织中可见HeLa、A549瘤组织内有明显的点状绿色荧光，而SKOV3、EC9706瘤组织内表达GFP的细胞数明显少于前两种瘤组织；HeLa、A549裸鼠移植瘤组织内大多数瘤细胞高表达CAR（卅），SKOV3、EC9706移植瘤组织内CAR表达水平较低（＋或-），表明瘤体内CAR表达水平与Ad．GFP的转导效率也呈正相关。结论：体内外实验均显示肿瘤细胞的CAR表达水平与5型腺病毒转导效率密切相关。肿瘤患者治疗前检测组织中CAR表达水平有助于规范腺病毒载体的基因治疗药物的个体化使用。

柯萨奇病毒-腺病毒受体在结肠癌组织中的表达及意义

目的:柯萨奇病毒-腺病毒受体(coxsackie and adenovirus receptor,CAR)介导腺病毒与靶细胞之间的粘附,是腺病毒转染靶细胞的限速因子。本文分析CAR在瘤旁上皮组织、结肠肿瘤原发灶和淋巴结转移灶中的表达,为设计腺病毒载体基因治疗方案提供依据。方法:采用免疫组织化学方法检测62例结肠癌组织中CAR的表达,分析CAR表达与临床病理特征的关系。结果:与瘤旁上皮组织相比,肿瘤组织普遍存在CAR表达下调,但CAR在原发灶和淋巴结转移灶的表达水平无明显差异。CAR的表达水平与患者年龄及肿瘤大小相关。结论:结肠癌组织中存在CAR表达下调,提示在设计腺病毒载体基因治疗方案时应充分考虑CAR表达变化对疗效的影响。

小细胞肺癌组织中柯萨奇-腺病毒受体阳性表达的意义

目的:探讨柯萨奇-腺病毒受体(CAR)在小细胞肺癌组织中阳性表达的意义。方法:应用免疫组化EnVision法检测病理诊断明确的31例小细胞肺癌组织中CAR的表达情况,用3例正常的肺组织做阴性对照。结果:31例小细胞肺癌组织全部呈阳性表达,阳性率为100%,3例非肿瘤性肺组织均呈阴性表达。结论:在小细胞肺癌组织中高表达CAR可能提示腺病毒为载体的基因治疗将在小细胞肺癌的治疗领域起重要的作用。

受体依赖性TRAIL重组腺病毒对人肺癌细胞凋亡的诱导作用

目的:观察重组TRAIL腺病毒(Ad5-TRAIL及Ad5F35-TRAIL)对人非小细胞肺癌(nonsmall-cell lung cancer,NSCLC)原代培养细胞和细胞株的凋亡诱导作用,探讨两种Ad-TRAIL用于肺癌基因治疗的价值。方法:采用流式细胞术检测人肺癌细胞系A549、Z793、QG56和NCI-H520及10例原代培养肺癌细胞中CAR和CD46的表达水平;分别以Ad5-TRAIL及Ad5F35-TRAIL重组腺病毒按MOI 10和50感染上述细胞,48 h后Annexin V-FITC双标法流式细胞术检测细胞的早期凋亡。结果:A549、Z793、QG56和NCI-H520 4株肺癌细胞中,CD46的表达均明显高于CAR表达。Z793、QG56细胞对Ad5-TRAIL和Ad5F35-TRAIL的作用较敏感,MOI=10感染后凋亡率分别为(11.76±2.10)%、(15.96±2.89)%和(6.05±1.58)%、(10.11±1.26)%,显著高于对照组[(2.33±0.37)%和(5.95±1.89)%,P<0.05];MOI=50感染时NCI-H520细胞凋亡率分别为(12.89±3.2)%和(9.08±1.35)%,与对照组(7.04±2.17)%相比差异无统计学意义(P>0.05);Ad5-TRAIL和Ad5F35-TRAIL均不能诱导A549细胞凋亡。10例原代肺癌细胞CD46表达也明显较CAR高;Ad5-TRAIL或Ad5F35-TRAIL感染后,5例的原代肺癌细胞检测到凋亡;与Ad5-TRAIL相比,Ad5F35-TRAIL诱导的凋亡率更高。结论:两种TRAIL重组腺病毒对非小细胞肺癌细胞均有凋亡诱导作用,Ad5F35-TRAIL的作用强于Ad5-TRAIL,更适合于非小细胞肺癌的基因治疗。

柯萨奇病毒-腺病毒受体在肺癌组织中的表达意义

目的:初步探讨柯萨奇病毒-腺病毒受体(Coxsackie and adenovirus receptor,CAR)与人肺鳞状细胞癌及腺癌发生、发展的关系,为设计腺病毒载体对肺癌进行基因治疗的方案提供理论依据。方法:采用免疫组织化学方法检测112例肺癌(包括鳞状细胞癌65例,腺癌47例)组织中CAR的表达情况,利用统计学方法(SPSS10.0软件)分析CAR的表达在鳞状细胞癌、腺癌以及癌旁正常肺组织中有否差异。结果:正常肺组织中仅有少量CAR的表达。两类型肺癌分别与癌旁组织CAR的表达差别有统计学意义(P<0.01),在鳞状细胞癌中CAR的表达较癌旁组织提高34.75%,在腺癌中CAR的表达较癌旁组织提高38.30%。其中,鳞状细胞癌的CAR阳性率为43.08%,腺癌的阳性率为70.21%,两类型肺癌间CAR表达的阳性率差别亦有统计学意义(P<0.01)。但CAR的表达在肺鳞状细胞癌不同组织学分级中(Ⅰ～Ⅲ级)的差别未有统计学意义(P>0.05)。此外,我们发现在肺腺癌中所包含的细支气管肺泡癌部分几乎都有CAR的表达。结论:肺癌组织(鳞状细胞癌和腺癌)CAR的表达普遍提高,提示CAR与肺癌的发生、发展有一定的联系,并且与肺腺癌的形成及发展的关系更为密切,因此为进一步实现用腺病毒载体对肺癌进行基因治疗提供了理论基础。

肺癌患者柯萨奇病毒和腺病毒受体的表达及其临床意义

目的:探索柯萨奇病毒和腺病毒受体(Coxsackie and adenovirns receptor,CAR)与肺癌的关系,为使用基于腺病毒载体的基因治疗方案提供依据方法:运用免疫组织化学法检测112例肺癌患者肿瘤组织、癌旁组织以及正常组织中CAR的表达;通过RT-PCR和Western blot法检测32例患者CARmRNA及CAR蛋白的表达。结果:在正常肺组织中未见CAR表达,癌旁组织中可见少量的CAR表达鳞状细胞癌和腺癌中CAR阳性表达比率分别为43%和70%,均较癌旁组织明显升高同时,腺癌中CAR的表达也显著高于鳞状细胞癌,RT-PCR和Western blot法进一步证实免疫组织化学法检测结果。结论:肺癌中CAR过表达,在腺癌中尤为明显,提示CAR可能参与了肺癌的发生发展。