上皮細胞激酶—2與層黏連蛋白5—γ—2在食管鱗癌細胞株TE13中相關性的研究

[摘要] 目的 探討上皮細胞激酶與層黏連蛋白laminin 5-γ-2在食管鱗癌細胞株TE13中表達相關性。 方法 采用細胞免疫組化法檢測EphA2與laminin 5-γ-2在食管鱗狀細胞癌細胞株TE13中的表達相關性；采用RT-PCR方法檢測EphA2與LN5-γ-2 mRNA表達的關系；采用Western Blot法檢測食管鱗狀細胞癌細胞株TE13中經RNA干擾技術特異性沉默EphA2后LN5-γ-2蛋白表達的變化。 結果 細胞免疫組化結果顯示LN5-γ-2蛋白在細胞胞漿表達，且沉默EphA2后，LN5-γ-2表達減少。RT-PCR及Western Blot實驗結果顯示在食管鱗狀細胞癌細胞株TE13中，LN5-γ-2 mRNA及蛋白均隨EphA2的沉默而受抑制（P<0.05）。 結論 LN5-γ-2表達受EphA2的調控，可能為EphA2的靶基因。EphA2可能通過調控laminin5-γ-2對食管鱗癌腫瘤血管生成擬態進行調節。

[關鍵詞] 食管鱗癌；上皮細胞激酶；層黏連蛋白5-γ-2

[中圖分類號] R735.1 [文獻標識碼] A [文章編號] 2095-0616（2012）21-39-03

The correlation of EphA2 and laminin 5-γ-2 in human esophageal cell TE13 line in vitro

ZHU Sumin1 FENG Yadong2 JIN Hailin3 SHI Ruihua2

1.Medical Center for Digestive Diseases，the Second Affiliated Hospital of Nanjing Medical University，Nanjing 210011，China； 2.Department of Digestion，the First Affiliated Hospital of NJMU，Nanjing 210029，China； 3.Gastrointestinal Endoscopy Center，Jiangsu Provincial Hospital of TCM，Nanjing 210029，China

[Abstract] Objective To investigate the correlation of EphA2 and laminin 5-γ-2 in human esophageal cell TE13 line in vitro. Methods In the present study，human esophageal squamous cell carcinoma TE13 lines were examined for LN-5 expression immunohistochemically，using an antibody against LN-5-γ-2 chain.TE13 cell line with scilencing EphA2 gene were examined for LN-5-γ- 2 chain levels by Western blotting and RT-PCR. Results Laminin 5-γ-2 could be detected in cytoplasm in TE13 and the level of expression of lamanin 5-γ-2 decreased with silencing EphA2 immunohistochemically.RT-PCR and Western blot showed that the expression of laminin 5-γ-2 in TE13 was inhibited with silencing EphA2（P<0.05）. Conclusion The expression of laminin 5-γ-2 positively correlates with expression of EphA2.Laminin 5-γ-2 may be a potential target gene of EphA2.

[Key words] Esophageal squamous cell carcinoma；Epithelial cell kinase；Laminin 5-γ-2

我國是食管癌的高發區，研究食管癌的發病機制、提高早期診斷率、尋找新的治療途徑是食管癌防治的關鍵[1]。食管癌是生長迅速的腫瘤，因此食管癌組織內常常存在著乏氧微環境。缺氧狀態可以刺激腫瘤細胞合成和分泌大量促血管生成因子如血管內皮生長因子（VEGF）等，促進腫瘤血管的生成，促進惡性腫瘤生長、浸潤和轉移。血管生成擬態（vasculogenic mimicry，VM）是近年來發現的一種存在于高侵襲性腫瘤的血液供應方式，是由腫瘤細胞通過自身變形和基質重塑產生血管樣通道，并與宿主血管相連，使腫瘤獲得足夠血供的一種血管化途徑。VM的存在對腫瘤浸潤、轉移有著巨大影響，與惡性腫瘤的預后密切相關[2-3]。層黏連蛋白（laminin，LN）是一種有多種生物學功能細胞外基質的糖蛋白，它們在基底膜的構建及細胞粘附、生長、遷移和分化中扮演著至關重要的角色[4]。……