【作者】 杨青；

【导师】 肖桂山； 尚东；

【作者基本信息】 大连理工大学 ， 化学工程（专业学位）， 2019， 硕士

【摘要】 口腔癌是头颈部较常见的恶性肿瘤之一,其发病率在全身常见恶性肿瘤中位列第六位。口腔癌患者的五年生存率为50%～60%,但I期临床疾病患者的五年生存率可达到80%以上,而晚期的口腔癌患者的生存率仅有20%,不幸的是三分之二的患者在癌症晚期才被临床诊断,此时癌症已经发生扩散和转移,难于治疗且治疗后预后极差。因此一方面需要通过早期发现,早期诊断和早期治疗来提高口腔癌患者的生存率,另一方面需要开发研究更具有靶向性的治疗方法来应对口腔癌的转移。OLA1是一个非常古老的、在进化中高度保守个的GTP酶,已有研究表明OLA1与肿瘤能量代谢、肿瘤细胞增殖、凋亡和迁移有着密切关系,深入研究OLA1与口腔癌的关系是一个潜在的抗癌策略。我们首先通过Western blot实验检测了OLA1在口腔正常角质细胞(Nhoks)和口腔癌细胞,包括口腔癌高转移细胞Um-1和Cal-27以及口腔原位癌细胞Um-2中的表达差异,实验结果发现,OLA1在口腔癌细胞中的表达显著低于口腔正常细胞,且随着口腔癌细胞转移程度的增加OLA1的表达逐渐减少。因此我们猜想OLA1在口腔癌中是一个抑癌基因,对口腔癌转移起到至关重要的作用。进一步通过在体外干扰OLA1的表达来确定OLA1对口腔癌转移的作用,划痕实验、Transwell侵袭和转移实验表明,敲低细胞中OLA1的表达可以促进口腔癌的转移和侵袭能力;反之,过表达OLA1可以抑制口腔癌的转移,同时,过表达OLA1可以降低口腔癌对紫杉醇的耐药性。为了进一步研究OLA1在口腔癌中的作用机制,我们通过qRT-PCR和Western blot实验探究OLA1的上下游机制。实验结果表明,OLA1可能是通过参与EMT途径和TGF-β信号通路来指导口腔癌的转移。本研究的结果表明,OLA1可能是一个潜在的治疗口腔癌转移的分子靶标,为探究口腔癌的发病机制提供了新的观点,同时为口腔癌的转移和预后提供了新的诊断和治疗策略。更多还原

【Abstract】 Oral cancer is one of the more common malignant tumors of the head and neck,ranking sixth in common malignant tumors.The five-year survival rate of patients with stage I clinical disease can reach more than 80%,while the survival rate of patients with advanced oral cancer is only 20%,unfortunately three Two-thirds of patients are clinically diagnosed in the late stages of cancer,when cancer has spread and metastasized,it is difficult to treat and the prognosis is very poor after treatment.Therefore,it is necessary to develop and research more targeted treatments to deal with the metastasis of oral cancer.OLA1 is a very old GTP enzyme that is highly conserved in evolution.Studies have shown that OLA1 is closely related to tumor energy metabolism,tumor cell proliferation,apoptosis and migration.It is a potential anti-cancer strategy to study the relationship between OLA1 and oral cancer.We first examined the expression difference of OLA1 in oral normal keratinocytes(Nhoks)and oral cancer cells,including oral metastatic cells Um-1 and Cal-27,and oral carcinoma cells Um-2 by Western blot.The results showed that the expression of OLA1 in oral cancer cells was significantly lower than that in normal oral cells,and the expression of OLA1 decreased with the increase of oral cancer cell metastasis.Therefore,we suspect that OLA1 is a tumor suppressor gene in oral cancer and plays a vital role in oral cancer metastasis.Furthermore,the effect of OLA1 on oral cancer metastasis was determined by interfering with the expression of OLA1 in vitro.Scratch test,Transwell invasion and metastasis experiments showed that knocking down expression of OLA1 in oral cancer cells can promote the metastasis and invasion;on the contrary,overexpression of OLA1 can inhibit the metastasis of oral cancer,and overexpression of OLA1 can reduce the resistance of oral cancer to paclitaxel.To further investigate the mechanism of action of OLA1 in oral cancer,we explored the upstream and downstream mechanisms of OLA1 by qRT-PCR and Western blot.The experimental results indicate that OLA1 may be involved in the metastasis of oral cancer by participating in the EMT pathway and the TGF-β signaling pathway.The results of this study suggest that OLA1 may be a potential molecular target for the treatment of oral cancer metastasis,providing a new perspective for exploring the pathogenesis of oral cancer,and providing new diagnostic and therapeutic strategies for the metastasis and prognosis of oral cancer.更多还原