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目的探索乙酰肝素酶短发夹RNA(HPA-shRNA)对乳腺癌细胞的抗肿瘤作用。方法根据RNA干扰(interfering RNA,RNAi)序列设计原则,以乳腺癌HPA基因为靶基因,构建shRNA重组慢病毒表达载体,通过Western-blot检测HPA-shRNA的敲降作用;通过CCK8实验观察HPA-shRNA对乳腺癌MDA-MB-231细胞生长增殖的影响;运用Transwell实验检测HPA-shRNA对乳腺癌MDA-MB-231侵袭能力的影响。结果经测序分析证实HPA-shRNA1重组慢病毒载体构建成功;在MDA-MB-231细胞内,HPA-shRNA1有效降低了HPA蛋白的表达水平(P<0.05);通过体外实验证明了HPA-shRNA1能够抑制乳腺癌MDA-MB-231细胞的生长增殖及侵袭能力。结论 HPA-shRNA1重组慢病毒载体可有效抑制HPA在乳腺癌细胞中的表达,并且证实HPA-shRNA对MDA-MB-231乳腺癌细胞具有良好的抗肿瘤作用。
Objective To explore the antitumor effect of heparanase short hairpin RNA (HPA-shRNA) on breast cancer cells. Methods According to the design principles of RNA interference (RNAi) sequence, HPA-shRNA expression vector was constructed by targeting HPA gene of breast cancer. The knockdown effect of HPA-shRNA was detected by Western-blot. The effect of HPA- shRNA on breast cancer MDA-MB-231 cells proliferation; Transwell assay using HPA-shRNA on breast cancer MDA-MB-231 invasion. Results HPA-shRNA1 recombinant lentiviral vector was constructed successfully by sequencing analysis. HPA-shRNA1 effectively reduced HPA protein expression in MDA-MB-231 cells (P <0.05). HPA-shRNA1 was able to Inhibit the growth, proliferation and invasion of breast cancer MDA-MB-231 cells. Conclusion HPA-shRNA1 recombinant lentiviral vector can effectively inhibit the expression of HPA in breast cancer cells, and confirmed HPA-shRNA has a good anti-tumor effect on MDA-MB-231 breast cancer cells.