人肺癌细胞的肿胀应激及其信号通路分析

Analysis of swelling stress and signaling pathway of human lung cancer cells

目的通过比较肺癌与对应癌旁组织的含水量、细胞核大小以及丝裂原激活蛋白激酶(MAPK)磷酸化激活情况以明确肺癌细胞是否存在肿胀应激。方法采用干湿质量比较法检测10例人肺腺癌、12例肺鳞癌以及对应的癌旁正常肺组织新鲜标本的含水量差别。用磷酸化和非磷酸化抗体结合免疫组织化学染色方法,检测30例肺腺癌、32例肺鳞癌、10例正常肺泡上皮细胞和10例正常肺支气管上皮细胞c-Jun氨基末端激酶(JNK)、胞外信号调节激酶1/2(ERK1/2)和p38 MAPK的表达及磷酸化激活情况。最后采用HE染色结合图像分析软件估算肺腺癌和肺鳞癌的癌细胞和对应癌旁正常肺泡上皮细胞及支气管上皮细胞的细胞核大小差别。结果干湿质量比较法发现肺鳞癌和肺腺癌的平均含水量均显著高于其对应的癌旁正常组织。免疫组织化学染色发现p38 MAPK在肺腺癌和肺鳞癌及癌旁组织均高表达。ERK在肺腺癌和鳞癌组织均弱表达,但是在肺癌旁组织高表达。JNK在肺腺癌及癌旁组织均弱表达。在肺泡上皮组织和支气管上皮组织中JNK、p38 MAPK、ERK1/2均未被磷酸化激活。但是,肺腺癌、肺鳞癌组织中JNK被磷酸化激活,而p38 MAPK及ERK1/2未被磷酸化激活。肺癌细胞的细胞核明显大于对应的正常肺上皮细胞的细胞核。结论肺癌细胞存在肿胀应激,不同组织类型肺癌细胞肿胀的信号通路存在差异。

Objective To investigate whether swelling stress exists in lung cancer cells by comparing water content and nucleus size as well as mitogen activated protein kinase(MAPK)phosphorylation activation between lung cancer tissues and corresponding paracancerous tissues.Methods Differences in water content between fresh specimens of 10 human lung adenocarcinoma,12 lung squamous cell carcinoma,and corresponding paracancerous normal lung tissues were examined through“dry and wet”mass comparison.The expressions and phosphorylation activation of mitogen activated protein kinases JNK,ERK1/2,and p38 in 30 lung adenocarcinoma samples,32 lung squamous cell carcinoma samples,and alveolar epithelial cells and lung bronchial epithelial cells of 10 paracancerous normal tissues samples were detected with their phosphorylated and non-phosphorylated antibodies and by immunohistochemical staining.Nucleus size differences between cancer cells from lung adenocarcinoma and lung squamous cell carcinoma samples and corresponding paracancerous normal lung alveolar epithelial cells or bronchial epithelial cells were estimated by HE staining and analyzed with image analysis software.Results The average water content of lung squamous cell carcinoma and lung adenocarcinoma samples were significantly higher than that of their corresponding paracancerous normal tissues.Immunohistochemistry showed that p38 was highly expressed in all samples of lung adenocarcinoma,lung squamous cell carcinoma,and paracancerous tissues.ERK was weakly expressed in lung adenocarcinoma and squamous cell carcinoma samples,but highly expressed inparacancerous tissues.JNK was weakly expressed in lung adenocarcinoma and paracancerous tissues.JNK,p38 MAPK,and ERK1/2 were not activated through phosphorylation in alveolar or bronchial epithelial tissues.However,JNK,not p38 or ERK1/2,was phosphorylated in lung adenocarcinoma and lung squamous cell carcinoma tissues.The average nucleus size of lung cancer cells was significantly larger than those of their corresponding paracancerous normal lung epithelial cells.Conclusion There is swelling stress in lung cancer cells,and different types of human lung cancer cells have different swelling stress signaling pathways.