宫颈癌细胞EGFR过表达对肿瘤细胞生物学行为的影响及机制研究

Effect of EGFR overexpression on biological behavior of cervical cancer cells and its mechanism

目的 探索表皮生长因子受体(epidermal growth Factor receptor,EGFR)过表达对宫颈癌细胞生长,侵袭及凋亡等生物学行为的影响,并探讨其潜在机制。方法 构建EGFR过表达、EGFR空载质粒和空白对照组的宫颈癌SiHa细胞,应用MTT检测、细胞划痕实验和Transwell实验分别检测EGFR过表达对SiHa细胞增殖、迁移和侵袭能力的影响,采用流式细胞技术分析细胞周期,并应用RT-PCR和westem blot检测PKM2和STAT3 mRNA和蛋白表达水平。结果 用慢病毒转染建立过表达EGFR的SiHa宫颈癌细胞后,明显促进了细胞的增殖、侵袭及迁移,且具有生物学统计意义,P <0.05;采用流式细胞技术分析细胞周期结果显示:EGFR过表达组、对照组和空载组三者中,EGFR高表达组DNA合成期细胞数量较其它两组显著升高,而合成前期细胞数量明显较其它两组减少,且P <0.05。EGFR过表达组中的PKM2、STAT3的表达量明显高于对照组和空载组。结论 EGFR表达增加显著促进宫颈癌细胞的增殖、侵袭、转移等恶性生物学行为,过表达EGFR的SiHa细胞生长失控,分裂活跃。宫颈癌中EGFR的表达与PKM2、STAT3表达呈正比,可能EGFR能通过PKM2-STAT3途径作用于宫颈癌SiHa细胞。

Objective To investigate the effect of epidermal growth factor receptor(EGFR)overexpression on the growth, invasion and apoptosis of cervical cancer cells and its underlying mechanism. Methods Si Ha cells with EGFR overexpression, Si Ha cells with EGFR empty vector and blank control Si Ha cells were constructed. The effects of EGFR overexpression on the proliferation, migration and invasion of Si Ha cells were detected by MTT assay, cell scratch assay and transwell assay. The cell cycle was analyzed by flow cytometry method, The m RNA and protein expression levels of PKM2 and STAT3 were detected by RT-PCR and Western blot assay. Results After transfection with lentivirus, Si Ha cervical cancer cells overexpressing EGFR significantly promoted cell proliferation, invasion and migration, and which had significant differences(P < 0.05). The results of cell cycle analysis by flow cytometry showed that the number of cells in DNA synthesis phase of EGFR overexpression group was significantly higher than that of the other two groups, while the number of cells in pre-synthesis phase of EGFR overexpression group was significantly lower than that of the other two groups(P < 0.05). The expression levels of PKM2 and STAT3 in EGFR overexpression group were significantly higher than those in control group and empty group. Conclusion EGFR overexpression significantly promoted the proliferation, invasion, metastasis and other malignant biological behaviors of cervical cancer cells. The expression of EGFR was positively correlated with the expression of PKM2 and STAT3 in cervical cancer. It is possible that EGFR can act on Si Ha cells through PKM2-STAT3 pathway.