蛋白酶活化受体2在葡萄膜黑色素瘤中的表达及对细胞增殖和侵袭的影响

目的：探讨蛋白酶活化受体2(protease-activated receptor 2，PAR2)在葡萄膜黑色素瘤(uveal melanoma，UM)中的表达，以及沉默人UM 细胞系M23中PAR2基因对细胞增殖和侵袭的影响。

方法：选取2012-02/2017-12在我院行手术治疗且资料完整的UM患者45例45眼，选取同期因眼部外伤行眼球摘除且葡萄膜正常的患者30例30眼，实时荧光定量PCR术检测UM和正常脉络膜组织中PAR2基因表达，培养M23细胞并分为PAR2干扰组、阴性对照序列组和空白组，实时荧光定量PCR技术检测细胞中PAR2基因表达，MTT法检测细胞增殖能力，Transwell法检测细胞迁移和侵袭能力。

结果：UM组织中PAR2 mRNA相对表达量为1.73±0.13，正常脉络膜组织中PAR2 mRNA相对表达量为1.06±0.10，差异有统计学意义(t=23.732，P<0.01)； UM组织中PAR2 mRNA相对表达量与病理学类型、巩膜浸润、视盘受累和眼外生长有关，差异有统计学意义(P<0.05)； PAR2干扰组细胞中PAR2 mRNA相对表达量低于阴性对照序列和空白组，差异有统计学意义(P<0.05)； PAR2干扰组细胞24、48、72和96h时吸光度A值低于阴性对照组和空白组，差异有统计学意义(P<0.05)； PAR2干扰组迁移细胞数和侵袭细胞数低于阴性对照序列组和空白组(P<0.05)。

结论：PAR2在UM组织中呈高表达，且与肿瘤高转移风险有关，特异性沉默M23细胞中PAR2基因表达可有效抑制细胞增殖、迁移及侵袭。

Expression of protease-activated receptor 2 in uveal melanoma and its effect on cell proliferation and invasion

AIM: To investigate the expression of protease-activated receptor 2(PAR2)in uveal melanoma(UM), and the effects of silencing the expression of PAR2 gene on proliferation and invasion of human UM cell line M23.

METHODS: A total of 45 patients(45 eyes)with UM who underwent surgical treatment with complete information in our hospital were selected from February 2012 to December 2017. In the same period, 30 patients(30 eyes)who underwent eyeball removal due to ocular trauma and most of the uvea were normal were selected. Real-time quantitative PCR was used to detect the expressions of PAR2 gene in UM and normal choroidal tissues. M23 cells were cultured and divided into PAR2 interference group, negative control sequence group and blank group. Real-time quantitative PCR was used to detect the expression of PAR2 gene in cells. MTT assay was used to detect cell proliferation, and transwell assay was used to detect cell migration and invasion.

RESULTS: The relative expression level of PAR2 mRNA was 1.73±0.13 in UM tissues, and 1.06±0.10 in normal choroid tissues(t=23.732, P<0.001). The relative expression level of PAR2 mRNA in UM tissues was associated with pathological type, scleral invasion, optic disc involvement and extraocular growth(P<0.05). The relative expression level of PAR2 mRNA in PAR2 interference group was lower than that in negative control sequence group and blank group(P<0.05). The absorbance A values at 24h, 48h, 72h and 96h in the PAR2 interference group cells were lower than those in negative control sequence group and blank group(P<0.05). The number of migrated cells and the number of invasive cells in PAR2 interference group were lower than those in negative control sequence group and blank group(P<0.05).

CONCLUSION: PAR2 was highly expressed in UM tissues and was associated with high risk of tumor metastasis. Specific silencing of PAR2 gene expression in M23 cells could effectively inhibit cell proliferation, migration and invasion.