[慢病毒介导的表皮生长因子样结构域7沉默对人喉癌Hep-2细胞增殖和侵袭的影响]
[Effects of lentivirus-mediated epidermal growth factor-like domain 7 silencing on proliferation and invasion of human laryngeal carcinoma Hep-2 cells].
作者信息
Li Jingjia, Ye Jin, Zhuang Shimin, Wang Tao, Wang Zhiyuan, Chang Lihong, Zhang Gehua
机构信息
Department of Otorhinolaryngology Head and Neck Surgery, Third Affiliated Hospital of SUN Yat-sen University, Guangzhou 510630, China.
Department of Otorhinolaryngology Head and Neck Surgery, Third Affiliated Hospital of SUN Yat-sen University, Guangzhou 510630, China. Email:
出版信息
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2014 Aug;49(8):643-8.
OBJECTIVE
To explore the effects of epidermal growth factor-like domain 7 (EGFL7) gene silencing on the proliferation and invasion ablity of laryngeal carcinoma cells.
METHODS
A lentiviral vector expressing EGFL7 shRNA was constructed and transfected into human laryngeal carcinoma Hep-2 cells. The expressions of EGFL7 mRNA and protein were detected by Real-time PCR and Western blot, respectively. Cell proliferation was evaluated by CCK-8 assay, cell cycle and apoptosis were tested by flow cytometry, and cell invasion was detected by transwell invasion assay.
RESULTS
The relative expression level s of EGFL7 mRNA and protein in EGFL7-SuRNA group were svgnificantly lower than control group (P < 0.001). Western blot analysis proved that the relative expression of EGFL7 protein in NC group, Lenti-NC group and Lenti-EGFL7 group was (0.39 ± 0.12),(0.36 ± 0.14) and (0.07 ± 0.04), respectively. EGFL7 expression in Lenri-EGFL7 group was significantly inhibited than NC group (P < 0.001), which confirmed that the recombinant lentivirus was successfully transfected into Hep-2 cells. The proliferation of Hep-2 cells was significantly inhibited after transfection (P < 0.01). Compared with the NC group and Lenti-NC group, the proportion of cells in S phase was significantly increased in Lenti-EGFL7 group (P < 0.01), and the proportion in G1 phase was significantly decreased (P < 0.05). Cell apoptosis assay showed that the apoptotic rate in Lenti-EGFL7 group (66.2 ± 1.28) % was significantly increased in NC group (6.09 ± 3.28) % and Lenti-NC group (9.86 ± 2.13) %. In Transwell invision assay, the mean number of cells coming through the Metrigel in Lenti-EGFL7 group was significantly decreased than in the NC group and Lenti-NC group (P < 0.01).
CONCLUSION
The proliferation and invasion ablity of laryngeal carcinoma Hep-2 cells can be inhibited by siRNA mediated EGFL7 gene silencing.
目的
探讨表皮生长因子样结构域7（EGFL7）基因沉默对喉癌细胞增殖和侵袭能力的影响。
方法
构建表达EGFL7 shRNA的慢病毒载体并转染人喉癌Hep-2细胞。分别采用实时荧光定量PCR和蛋白质印迹法检测EGFL7 mRNA和蛋白的表达。采用CCK-8法评估细胞增殖，通过流式细胞术检测细胞周期和凋亡，采用Transwell侵袭实验检测细胞侵袭能力。
结果
EGFL7-siRNA组EGFL7 mRNA和蛋白的相对表达水平均显著低于对照组（P < 0.001）。蛋白质印迹分析表明，NC组、Lenti-NC组和Lenti-EGFL7组EGFL7蛋白的相对表达量分别为（0.39±0.12）、（0.36±0.14）和（0.07±0.04）。Lenti-EGFL7组EGFL7表达较NC组显著抑制（P < 0.001），证实重组慢病毒成功转染至Hep-2细胞。转染后Hep-2细胞的增殖受到显著抑制（P < 0.01）。与NC组和Lenti-NC组相比，Lenti-EGFL7组S期细胞比例显著增加（P < 0.01），G1期细胞比例显著降低（P < 0.05）。细胞凋亡检测显示，Lenti-EGFL7组凋亡率（66.2±1.28）%显著高于NC组（6.09±3.28）%和Lenti-NC组（9.86±2.13）%。在Transwell侵袭实验中，Lenti-EGFL7组穿过基质胶的细胞平均数显著低于NC组和Lenti-NC组（P < 0.01）。
结论
siRNA介导的EGFL7基因沉默可抑制喉癌Hep-2细胞的增殖和侵袭能力。
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