RNAi沉默GALNT7基因对宫颈癌细胞恶性行为的影响

摘要

Objective To investigate the influence of polypeptide N-acetylgalactosaminyltransferase 7 (GALNT7) gene to growth, invasiveness and migration of cervical cancer cells. Methods GALNT7 gene was silenced by RNA interference, took non-specificity sequence pSilencer 2.1 as negative control. After transfection of cervical cancer cells (HeLa and C33A cells), Western Blot was used to detect protein expres-sion of GALNT7 gene, the growth ability was detected by MTT assay, the ability to form colonies was evaluated by the colony formation assay, the invasion and migration ability were detected by matrigel invasion assay. Re-sults In experimental group the protein expression of GALNT7 was decreased significantly than that of control group, and the differences all had statistical significance(Pall<0.05). MTT assay revealed that cell growth was inhibited, the inhibitory rates were decreased by19% in HeLa cells and 22% in C33A cells, and compared with control group, the differences all had statistical significance (Pall<0.05). The ability to form colonies in experimental group were inhibited by 56%in HeLa cells and 52%in C33A cells than that of control group , and the differences all had statistical significance(Pall<0.05). In experimental group, the inhibitory rates on inva-sion were decreased by 48%in HeLa cells and 54%in C33A cells , and the inhibitory rate on migration was decreased by 30%in HeLa cells, and compared with control group, the differences all had statistical signifi-cance (Pall<0.05). Conclusion RNAi mediated GALNT7 down-regulation can inhibit the growth, invasion and migration of cervical cancer, and can be an important target to cervical cancer gene-therapies.%目的：探讨多肽N-乙酰氨基半乳糖转移酶7（polypeptide N-acetylgalactosaminyltransfer-ase 7， GALNT7）基因对宫颈癌细胞生长、侵袭及迁移能力的影响。方法利用RNA干扰技术沉默GALNT7（siR-GALNT7），以非特异性序列（pSilencer 2．1）转染细胞作为阴性对照，转染宫颈癌细胞HeLa和C33A细胞，采用MTT实验、集落形成实验、Transwell侵袭实验以及细胞划痕实验来检测干扰GAL-NT7基因后宫颈癌细胞生长、侵袭以及转移能力的变化。结果 Western Blot检测结果显示，转染siR-GALNT7质粒的实验组中两种宫颈癌细胞中GALNT7的蛋白表达水平明显降低，与转染siR-Ctrl的对照组比较，差异均有统计学意义（P均＜0．05）；siR-GALNT7使HeLa和C33A细胞生长活性分别下降了19％和22％，与对照组比较，差异均有统计学意义（P均＜0．05）；siR-GALNT7使HeLa和C33A细胞的集落形成能力分别下降了56％和52％，与对照组比较，差异均有统计学意义（P均＜0．05）；siR-GALNT7使HeLa和C33A细胞侵袭能力分别下降了48％和54％，与对照组比较，差异均有统计学意义（P均＜0．05）；siR-GALNT7使HeLa细胞迁移能力下降了约30％，与对照组比较差异有统计学意义（P＜0．05）。结论 RNA干扰在体外明显降低了宫颈癌细胞中GALNT7基因的表达，同时抑制了宫颈癌细胞的生长侵袭以及迁移能力，GALNT7基因有可能成为宫颈癌基因治疗的重要靶点。