CAJ | 학술논문

目的：探讨转化生长因子(TGF)及表皮生长因子(EGF)信号对溶血磷脂酸(LPA)诱导口腔鳞癌细胞增殖的影响。方法：口腔鳞癌SAS细胞接种于96孔板中，以含不同浓度(0、1、5和10μmol/L)LPA的无血清DMEM分别处理细胞24和48 h后采用CCK-8法检测细胞增殖；免疫印迹检测10μmol/L LPA分别处理细胞0、0.5、2和4 h后对TGF-β下游信号分子Smad2的影响；应用不同浓度(10、20和50μg/mL)的TGF-β阻断抗体1D11和100 nmol/L EGFR阻断剂AG1478分别预孵育细胞2和4 h后，再加入LPA诱导，检测SAS细胞增殖情况。结果：LPA呈剂量效应和时间效应诱导SAS细胞增殖，LPA浓度增加到10μmol/L时SAS细胞数是对照组的近1.7倍(P<0.01)，细胞在LPA处理24 h后增殖速率较对照组增加22%(<0.05)。LPA可活化TGF-β，但TGF-β阻断抗体1D11并不能阻断LPA诱导的SAS增殖；相反，EGFR阻断剂AG1478能够抑制LPA诱导的SAS增殖，抑制率达到近77%。结论：LPA诱导的口腔鳞癌SAS细胞的增殖与EGFR活化信号有关，而与LPA诱导的TGF-β活化无关。P
목적：탐토전화생장인자(TGF)급표피생장인자(EGF)신호대용혈린지산(LPA)유도구강린암세포증식적영향。방법：구강린암SAS세포접충우96공판중，이함불동농도(0、1、5화10μmol/L)LPA적무혈청DMEM분별처리세포24화48 h후채용CCK-8법검측세포증식；면역인적검측10μmol/L LPA분별처리세포0、0.5、2화4 h후대TGF-β하유신호분자Smad2적영향；응용불동농도(10、20화50μg/mL)적TGF-β조단항체1D11화100 nmol/L EGFR조단제AG1478분별예부육세포2화4 h후，재가입LPA유도，검측SAS세포증식정황。결과：LPA정제량효응화시간효응유도SAS세포증식，LPA농도증가도10μmol/L시SAS세포수시대조조적근1.7배(P<0.01)，세포재LPA처리24 h후증식속솔교대조조증가22%(<0.05)。LPA가활화TGF-β，단TGF-β조단항체1D11병불능조단LPA유도적SAS증식；상반，EGFR조단제AG1478능구억제LPA유도적SAS증식，억제솔체도근77%。결론：LPA유도적구강린암SAS세포적증식여EGFR활화신호유관，이여LPA유도적TGF-β활화무관。P
OBJECTIVE:To investigate the effects of transforming growth factor-β(TGF-β) activation and epidermal growth factor receptor(EGFR) transactivation on lysophosphatidic acid (LPA)-induced human oral squamous cell carcinoma (OSCC) SAS cell proliferation. METHODS:SAS cells were seeded in 96-well plates and treated with LPA (0,1,5 and 10μmol/L) for 24 h and 48 h,and CCK-8 method was used to detect cell proliferation;Western blotting was used to detect the effect of LPA(10 μmol/L) on TGF-β activation by examining the expression of p-Smad2. Pre-incubation of SAS cells with TGF-βblocking antibody 1D11 (10,20 and 50μg/mL) and EGFR inhibitor AG1478 (100μmol/L) for 2 and 4 h were used to measure the effects of TGF-βactivation and EGFR transactivation on LPA-induced SAS proliferation. RESULTS:LPA stimulated SAS cell proliferation in dose- and time-dependent manners. The cell number in LPA(10μmol/L) treated group was 1.7 times higher than the number in untreated group (P<0.01). After 24 h incubation,the proliferation rate of LPA (10 μmol/L)treated group increased by 22% compared with the control group (P<0.05). LPA activated TGF-β. However,TGF-β blocking antibody 1D11 could not block LPA-induced SAS cell proliferation. In contrast,EGFR inhibitor AG1478 blocked LPA-induced SAS cell proliferation by 77%. CONCLUSION:LPA-induced SAS cell proliferation was dependent on EGFR transactivation and independent of TGF-βactivation.