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目的非肌性肌球蛋白重链9基因(non-muscular myosin heavy chain 9,MYH-9)在多种恶性肿瘤中高表达,且预后差。检测胃癌组织MYH-9的表达及利用RNA干扰(RNA interference,RNAi)技术特异性,抑制胃癌SGC-7901细胞中MYH-9的表达对细胞侵袭和转移的影响,探讨在胃癌治疗中将MYH-9作为基因治疗靶点的价值。方法选取2009-06-01-2010-01-01南昌大学第一附属医院126例胃癌及癌旁组织。利用RNAi技术将MYH-9siRNA转染到胃癌SGC-7901细胞中,蛋白质印迹法检测MYH-9蛋白的表达,RT-PCR法检测MYH-9mRNA的表达;运用细胞迁移和侵袭实验检测MYH-9siRNA下调胃癌SGC-7901细胞中MYH-9的表达对细胞侵袭转移的影响。结果 MYH-9在胃癌组织中的表达显著高于癌旁组织,χ2=77.83,P<0.001。胃癌MYH-9蛋白表达与肿瘤Lauren分型(χ2=5.296,P=0.020)、肿瘤分化程度(χ2=14.39,P=0.002)、肿瘤浸润深度(χ2=5.296,P=0.026)、肿瘤淋巴结转移数(χ2=11.15,P=0.003)、脉管癌栓(χ2=5.255,P=0.022)、肿瘤远处转移状态(χ2=6.683,P=0.009)和TNM分期(χ2=10.73,P=0.002)密切相关。MYH-9蛋白表达(HR=2.825,95%CI=1.104~3.706,P=0.041)、胃癌远处转移状态(HR=1.749,95%CI=0.983~3.503,P=0.035)和胃癌淋巴结转移数(HR=3.235,95%CI=1.579~5.358,P<0.001)可作为预测患者不良预后的独立指标。与空白对照组及阴性对照组比较,特异性siRNA可以高效抑制SGC-7901细胞MYH-9基因的表达,在mRNA水平其表达抑制率分别为87.5%(t=48.22,P<0.001)和86.9%(t=57.30,P<0.001);在蛋白质水平其表达抑制率分别为52.9%(t=11.01,P<0.001)和60.1%(t=15.53,P<0.001)。在侵袭实验中,实验组细胞数为(49.5±13.8)个,显著低于阴性对照组(112±4.34)个(t=15.53,P<0.001)和空白对照组(116±5.57)个(t=28.12,P<0.001)。在迁移实验中,实验组细胞数为(81.5±7.26)个,显著低于阴性对照组(185±7.33)个(t=47.29,P<0.001)和空白对照组(189±3.46)个,t=46.40,P<0.001。结论 MYH-9高表达患者预后较差,MYH-9可能成为胃癌基因治疗的新靶点。
Objective Non-muscular myosin heavy chain 9 (MYH-9) is highly expressed in many malignant tumors with poor prognosis. To detect the expression of MYH-9 in gastric cancer tissue and to detect the expression of MYH-9 in gastric cancer SGC-7901 cells by using the specificity of RNA interference (RNAi) technology to investigate the influence of MYH-9 on cell invasion and metastasis, 9 as a gene therapy target value. Methods Select 126 cases of gastric cancer and adjacent tissue from the First Affiliated Hospital of Nanchang University from June 2009 to January 2010. MYH-9 siRNA was transfected into gastric cancer cell line SGC-7901 by RNAi technique. The expression of MYH-9 protein was detected by Western blotting and MYH-9 mRNA was detected by RT-PCR. MYH-9 siRNA was detected by cell migration and invasion assay Effect of MYH-9 Expression in Invasion and Metastasis of Gastric Cancer SGC-7901 Cells. Results The expression of MYH-9 in gastric cancer tissues was significantly higher than that in adjacent tissues (χ2 = 77.83, P <0.001). The expression of MYH-9 in gastric cancer was correlated with Lauren’s classification (χ2 = 5.296, P = 0.020), tumor differentiation (χ2 = 14.39, P = 0.002), depth of tumor invasion (χ2 = 5.296, (Χ2 = 11.15, P = 0.003), vascular tumor thrombus (χ2 = 5.255, P = 0.022), tumor distant metastasis (χ2 = 6.683, )closely related. (HR = 2.825,95% CI = 1.104-3.606, P = 0.041), distant metastasis of gastric cancer (HR = 1.749,95% CI = 0.983-3.503, P = 0.035) and lymph node metastasis (HR = 3.235, 95% CI = 1.579-5.358, P <0.001) could be used as independent predictors of adverse prognosis. Compared with the blank control group and the negative control group, specific siRNA could effectively inhibit the MYH-9 gene expression in SGC-7901 cells, and the inhibitory rates were 87.5% (t = 48.22, P <0.001) and 86.9% (t = 57.30, P <0.001). The inhibition rates at the protein level were 52.9% (t = 11.01, P <0.001) and 60.1% (t = 15.53, P <0.001). In the invasion experiment, the number of cells in the experimental group was (49.5 ± 13.8), which was significantly lower than that in the negative control group (112 ± 4.34) (t = 15.53, P <0.001) and the blank control group (116 ± 5.57) = 28.12, P <0.001). In migration experiments, the number of cells in the experimental group was (81.5 ± 7.26), which was significantly lower than that in the negative control group (185 ± 7.33) (t = 47.29, P <0.001) and the blank control group (189 ± 3.46) = 46.40, P <0.001. Conclusion The prognosis of patients with MYH-9 overexpression is poor. MYH-9 may be a new target for gene therapy of gastric cancer.