目的 探討胃癌組織中人表皮生長因子受體(EGFR)、表皮生長因子受體-2(HER-2)和環氧合酶-2(COX-2)的表達及與臨床病理特征的關系。 方法 應用免疫組織化學Envision二步法,檢測70例胃癌組織中EGFR、HER-2和COX-2的表達情況,并結合其臨床病理特點進行分析。 結果 EGFR、HER-2和COX-2在胃癌組織中的表達陽性率分別為35.7%(25/70)、27.1%(19/70)、67.1%(47/70)。陽性表達與腫瘤分化程度、侵襲深度、有無淋巴結轉移及TNM分期有關(P<0.05),而與患者的性別及年齡、腫瘤部位和大小無關(Pgt;0.05)。EGFR、HER-2和COX-2三者之間在胃癌組織中的表達均呈正相關(P<0.05)。 結論 EGFR、HER-2和COX-2的表達參與胃癌的生長、侵襲和轉移過程。它們的聯合檢測有助于胃癌患者靶向藥物的選擇,也為胃癌的預后判斷提供客觀的參考指標。Objective To observe the expressions of epidermal growth factor receptor (EGFR), HER-2 and cyclooxygenase-2 (COX-2) in gastric carcinoma (GC) and to explore the relationship among them. Methods The envision immunohistochemical stain method was used to detect EGFR, HER-2 and COX-2 protein expressions in sample of 70 GC tissues. And their corresponding pathologic features were analyzed. Results The positive expression rates of EGFR, HER-2 and COX-2 protein in GC tissue were 35.7% (25/70), 27.1% (19/70) and 67.1% (47/70), respectively. The positive expression rates were closely relevant to the differentiation of the cancer, invasion depth, lymphatic metastasis and TNM (P<0.05), but not to the patient’ s sex, age, tumor site and size (P>0. 05). There was a stable positive correlation among EGFR, HER-2 and COX-2 expressions in GC tissues, respectively. Conclusions EGFR, HER-2 and COX-2 expressions participate in the development, invasion and metastasis process of GC. Combined detection can be regarded as an important symbol for guiding the molecular targeting therapy of GC, and judging the prognosis of GC.
目的 構建人flag-AWP1(associated with protein kinase C related kinase 1)基因的重組腺病毒載體作為AWP1的轉基因工具,研究AWP1在人血管內皮細胞ECV304中的表達與定位。方法 將flag-AWP1基因亞克隆到腺病毒穿梭載體pAdTrack-CMV,經酶切鑒定正確后PmeⅠ線性化,轉化至帶有骨架質粒pAdEasy-1的大腸桿菌BJ5183中進行重組; 經PacⅠ酶切鑒定和 PCR鑒定、線性化后,用Lipofectamine轉染293細胞進行包裝、反復感染以獲取高滴度重組病毒上清。再用獲得的重組腺病毒Ad-flag-AWP1感染ECV304細胞,激光掃描共焦顯微鏡下觀察AWP1在ECV304細胞中的表達與定位。結果 包裝病毒顆粒的PCR分析顯示,成功獲得了重組腺病毒Ad-flag-AWP1,該重組腺病毒能夠高效感染ECV304細胞,并在激光掃描共焦顯微鏡下觀察到AWP1在細胞內成功表達,且定位于細胞膜內側。結論 成功構建了flag-AWP1的重組腺病毒基因轉染載體,AWP1在ECV304細胞中的表達與定位提示AWP1可能具有細胞信號轉導因子的潛能,尚需進一步研究。