高危型HPV阳性宫颈癌细胞中YAP1与Wnt/β⁃catenin信号通路的关系及生物学意义

Relationship Between YAP1 and the Wnt/β⁃catenin Signaling Pathway in High⁃risk HPV⁃positive Cervical Cancer Cells and Its Biological Importance

高危型人乳头瘤病毒(HPV)感染是宫颈癌发病的明确危险因素,但相关的分子机制尚不清楚。YAP1是Hippo/YAP通路的关键分子,通过激活Wnt/β⁃catenin通路促进胃癌细胞、肝癌细胞增殖。为了阐明YAP1与Wnt/β⁃catenin信号通路在高危型HPV感染导致宫颈癌发病中的作用,本研究分析了高危型HPV阳性宫颈癌细胞中YAP1与Wnt/β⁃catenin信号通路的关系及生物学意义。首先检测高危型HPV阳性与阴性宫颈癌组织、高危型HPV阳性及阴性宫颈癌细胞与正常宫颈上皮细胞中YAP1、Wnt1、Wnt3a及β⁃catenin的表达,分析高危型HPV阳性宫颈癌组织中YAP1与Wnt1、Wnt3a、β⁃catenin的相关性;然后培养HPV16阳性的SiHa细胞及HPV18阳性的HeLa细胞,转染阴性对照(NC)或YAP1 siRNA、给予溶剂DMSO或Wnt/β⁃catenin激动剂SKL2001进行干预,检测细胞增殖活力A490及Wnt1、Wnt3a、β⁃catenin的表达水平。结果显示,高危型HPV阳性宫颈癌组织中YAP1、Wnt1、Wnt3a、β⁃catenin的表达水平高于高危型HPV阴性宫颈癌组织(P<0.05)且YAP1与Wnt1、Wnt3a、β⁃catenin呈正相关;转染YAP1 siRNA后,si⁃YAP1组SiHa细胞及HeLa细胞的A490水平及YAP1、Wnt1、Wnt3a、β⁃catenin表达水平均低于si⁃NC组(P<0.05);转染YAP1 siRNA的同时使用SKL2001,si⁃YAP1+SKL2001组A490水平、β⁃catenin表达水平均高于si⁃YAP1+DMSO组(P<0.05)。以上结果表明高危型HPV阳性的宫颈癌细胞中YAP1表达增加通过Wnt/β⁃catenin通路调控细胞增殖。

High⁃risk human papillomavirus(HPV)infection is a risk factor for cervical cancer,but the related molecular mechanism is not clear.YAP1 is a key molecule of the Hippo/YAP pathway.YAP1 promotes the proliferation of gastric cancer cells and liver cancer cells by activating the wingless type(Wnt)/β⁃catenin pathway.We wished to clarify the role of the YAP1 and Wnt/β⁃catenin signaling pathway in the pathogenesis of cervical cancer caused by high⁃risk HPV infection.We analyzed the relationship and biological importance of YAP1 and the Wnt/β⁃catenin pathway in high⁃risk HPV⁃positive cervical cancer cells.First,expression of YAP1,Wnt1,and Wnt3aβ⁃catenin in high⁃risk HPV⁃positive and⁃negative cervical cancer tissues,high⁃risk HPV⁃positive and⁃negative cervical cancer cells,and normal cervical epithelial cells was measured.Then,HPV16⁃positive SiHa cells and HPV18⁃positive HeLa cells were cultured and transfected with negative control(NC)or YAP1 siRNA,or given the solvent dimethyl sulfoxide(DMSO)or an agonist of Wnt/β⁃catenin(SKL2001).The cell⁃proliferation activity at 490 nm(A490)and expression of Wnt1 and Wnt3aβ⁃catenin were measured.Expression of YAP1,Wnt1,and Wnt3aβ⁃catenin in high⁃risk HPV⁃positive cervical cancer was higher than that in high⁃risk HPV⁃negative cervical cancer(P<0.05)and YAP1 was positively correlated with expression of Wnt1 and Wnt3aβ⁃catenin.After transfection with YAP1 siRNA,we discovered that A490 and expression of YAP1,Wnt1,and Wnt3aβ⁃catenin in SiHa cells and HeLa cells of the si⁃YAP1 group were lower than those of the si⁃NC group(P<0.05).SKL2001 was used while transfecting YAP1 siRNA,and A490 and expression ofβ⁃catenin in SiHa cells and HeLa cells of the si⁃YAP1+SKL2001 group were higher than those in the si⁃YAP1+DMSO group(P<0.05).Overall,our results showed that high expression of YAP1 in high⁃risk HPV⁃positive cervical cancer cells regulated cell proliferation through the Wnt/β⁃catenin pathway.