透明质酸、透明质酸酶和RGD抑制胃癌细胞SGC7901黏附侵袭的实验研究

Inhibitory effect of hyaluronic acid, hyaluronidase and RGD on adhesion and invasion of human gastric cancer cell line SGC7901

目的 探讨单独及联合应用透明质酸 (HA)、透明质酸酶 (Hase)和RGD对SGC790 1细胞黏附和侵袭细胞外基质 (ECM)的抑制作用。方法 采用间接免疫荧光法测定SGC790 1细胞表面CD4 4与integrinβ1蛋白表达。单独或联合应用HA、Hase和RGD处理体外培养的胃癌细胞SGC790 1;MTT法和Boyden小室模型分别测定其黏附力和侵袭力 ,同时观察细胞形态。结果 人胃癌细胞系SGC790 1表达CD4 4与integrinβ1蛋白。与对照组比较 ,Hase及RGD肽均明显抑制SGC790 1的黏附力(P <0 .0 0 1)和侵袭力 (P <0 .0 5 ) ,效果优于HA(P <0 .0 5 ) ;Hase +HA或三者联合的抑制效果优于任何单一抑制效果 (P <0 .0 0 1)。形态学观察发现 ,未处理组的部分细胞已经开始铺展 ,穿过人工基底膜后 ,表现出纤维母细胞样形态及形态各异的伪足 ;而处理组细胞形态较圆 ,伪足数目相对较少。结论 人胃癌细胞系SGC790 1表达功能性CD4 4和integrinβ1蛋白。单独及联合应用HA、Hase和RGD均不同程度抑制SGC790 1细胞对ECM的黏附和侵袭 ,以联合三者阻断效果为优。

Objective To investigate the inhibitory effect of hyaluronic acid (HA), hyaluronidase (Hase) and arg-gly-asp tripeptide (RGD) on adhesion and invasion of human gastric cancer cell line SGC7901. MethodsExpression of CD44 and integrin β1 protein on cell surface was determined by indirect fluorescence. After SGC7901 cells were treated with HA, Hase and RGD alone or in various combinations, their adhesion and invasion to ECM were measured by MTT and Boyden chamber method. Cell morphology was also observed. Results Expression of CD44 and integrin β1 protein on cell surface was detected in human gastric cancer cell line SGC7901. Hase or RGD alone could block the adhesion and invasion of SGC7901 cells to ECM in contrast to the control (P<0.001, P<0.05); their blocking effect was stronger than HA (P<0.05). The inhibitory effect of Hase + HA or a combination of the three agents was stronger than any single agent (P<0.001). Morphologically, the untreated cells adhered onto the matrigel had spread out presenting a fibroblast feature with variously shaped pseudopods, while the treated ones were kept round with relatively fewer pseudopods. ConclusionHA, Hase and RGD can inhibit the adhesion and invasion of SGC7901 cells expressing functional CD44 and integrin β1 protein to ECM, and a combination of the three agents may achieve the best inhibitory effect.