Sang Jianzhong, Liu Li, Tian Fang, Jin Hongjun, Yuan Linlin, Lou Weihua
Department of Otorhinolaryngology Head and Neck Surgery, the First Affiliated Hospital, Zhengzhou University, Zhengzhou, 450052, China.
Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2011 Aug;25(15):695-700.
To detect the expression of c-myc in the tissue of laryngeal squamous cell carcinoma. RNA interference(RNAi) was employed to inhibit the expression of c-myc in Hep-2 cells and to evaluate the effects of c-myc as a target for gene therapy in laryngeal carcinoma.
Immunohistochemistry was used to determine the protein levels of c-myc and Rb in 80 cases of laryngeal squamous cell carcinoma and 30 cases of polyp of vocal cord. Hep-2 cells were transfected with c-myc siRNA, c-myc protein and mRNA levels were detected using Western Blotting and RT-PCR. Cell viability was detected by MTT after the Hep-2 cells were transfected with c-myc siRNA for different times or transfected with different concentrations c-myc siRNA. The sensitivity of Hep-2 cells to 5-Fu transfected with or without c-myc siRNA was evaluated also by MTT. Hep-2 cells were transfected with c-myc siRNA in combination with 5-Fu for 48 h and then analyzed cell apoptosis by flow cytometry.
Immunohistochemical analysis showed that c-myc was highly expressed in the tissues of laryngeal squamous cell carcinoma while the expression of Rb was lower. The protein and mRNA levels of c-myc decreased after transfected with c-myc siRNA. The results of MTT showed that the c-myc siRNA inhibited Hep-2 cells growth in a concentration-dependent manner. When transfected with c-myc siRNA(50 nmol/L), the cells were inhibited in a time-dependent manner. Compared with the untransfected cells, the viability of transfected Hep-2 cells was significantly suppressed at the same concentration of 5-Fu (P < 0.05). C-myc siRNA combination with 5-Fu could obviously increase cell apoptosis, even in the low concentration of 5-Fu (P < 0.05).
The protein level of C-myc has highly expressed in tumor tissues. C-myc siRNA can effectively inhibit the expression of c-myc and has anti-proliferation effects, increasing the sensitivity of Hep-2 cells to 5-Fu. Therefore,c-myc might be a good target for cancer treatment.
检测c-myc在喉鳞状细胞癌组织中的表达。采用RNA干扰（RNAi）技术抑制Hep-2细胞中c-myc的表达，评估以c-myc为靶点进行喉癌基因治疗的效果。
采用免疫组织化学法检测80例喉鳞状细胞癌组织及30例声带息肉组织中c-myc和Rb的蛋白水平。用c-myc siRNA转染Hep-2细胞，采用蛋白质印迹法和逆转录聚合酶链反应检测c-myc蛋白和mRNA水平。用c-myc siRNA转染Hep-2细胞不同时间或转染不同浓度的c-myc siRNA后，采用噻唑蓝比色法检测细胞活力。还用噻唑蓝比色法评估转染或未转染c-myc siRNA的Hep-2细胞对5-氟尿嘧啶的敏感性。将c-myc siRNA与5-氟尿嘧啶联合转染Hep-2细胞48小时，然后通过流式细胞术分析细胞凋亡情况。
免疫组织化学分析显示，c-myc在喉鳞状细胞癌组织中高表达，而Rb表达较低。用c-myc siRNA转染后，c-myc的蛋白和mRNA水平降低。噻唑蓝比色法结果显示，c-myc siRNA以浓度依赖的方式抑制Hep-2细胞生长。当用c-myc siRNA（50 nmol/L）转染时，细胞以时间依赖的方式受到抑制。与未转染细胞相比，相同浓度的5-氟尿嘧啶作用下，转染的Hep-2细胞活力明显受到抑制（P<0.05）。c-myc siRNA与5-氟尿嘧啶联合使用可明显增加细胞凋亡，即使在低浓度5-氟尿嘧啶时也是如此（P<0.05）。
C-myc蛋白水平在肿瘤组织中高表达。C-myc siRNA可有效抑制c-myc的表达，具有抗增殖作用，增加Hep-2细胞对5-氟尿嘧啶的敏感性。因此，c-myc可能是癌症治疗的一个良好靶点。
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