本文選題：SLC25A10 + 肝癌　； 參考：《大連醫(yī)科大學(xué)》2017年碩士論文

【摘要】：世界范圍內(nèi),肝癌作為一種常見腫瘤,其發(fā)病率和死亡率均居高不下。有數(shù)據(jù)顯示,全球50%的新發(fā)和死亡肝癌病例均來(lái)自中國(guó)。世界衛(wèi)生組織預(yù)計(jì),如不采取緊急行動(dòng),2015年至2030年間中國(guó)將有約1000萬(wàn)人死于肝癌。目前,手術(shù)仍作為肝癌治療的主要手段,但手術(shù)后患者的五年生存率仍僅為15%-40%。因此,尋找新的治療方法和新的治療靶點(diǎn)是目前肝癌臨床的當(dāng)務(wù)之急,也是本課題的研究重點(diǎn)。人溶質(zhì)載體蛋白第25號(hào)家族中A亞家族的第10位成員SLC25A10作為一種線粒體膜上的轉(zhuǎn)運(yùn)蛋白,主要將蘋果酸和琥珀酸等二羧酸鹽從線粒體內(nèi)轉(zhuǎn)運(yùn)至線粒體外,以交換磷酸鹽、硫酸鹽及硫代硫酸鹽,從而為糖異生和尿素合成等過(guò)程提供底物,進(jìn)而維持TCA循環(huán)過(guò)程中的中間產(chǎn)物在線粒體內(nèi)外的分布和穩(wěn)態(tài)。近年來(lái),有多篇文獻(xiàn)報(bào)道轉(zhuǎn)運(yùn)蛋白SLC25A10在多種腫瘤中表達(dá)水平較高,但其在肝癌中的表達(dá)水平如何,鮮有文獻(xiàn)報(bào)道,且其分子機(jī)制尚不明確。根據(jù)前期實(shí)驗(yàn)結(jié)果證實(shí),轉(zhuǎn)運(yùn)蛋白SLC25A10在肝癌中有較高水平的表達(dá)且能夠抑制肝癌凋亡的發(fā)生,本課題旨在進(jìn)一步明確SLC25A10抑制肝癌凋亡發(fā)生的分子機(jī)制,找到治療肝癌的新的潛在的靶點(diǎn),更好的對(duì)肝癌進(jìn)行靶向治療。通過(guò)對(duì)多例臨床肝癌組織和癌旁組織進(jìn)行免疫組化分析,結(jié)果顯示,SLC25A10于肝癌組織中高表達(dá)。同時(shí),通過(guò)Realtime-PCR和Western Blot,在mRNA和蛋白水平對(duì)以上結(jié)果進(jìn)行了進(jìn)一步驗(yàn)證,得到了同樣的結(jié)果。然后,通過(guò)慢病毒感染的方式,分別對(duì)SLC25A10表達(dá)水平較低的肝癌細(xì)胞SMMC-7721進(jìn)行SLC25A10的穩(wěn)定高表達(dá),對(duì)SLC25A10表達(dá)水平較高的肝癌細(xì)胞HepG2進(jìn)行SLC25A10的穩(wěn)定低表達(dá)。通過(guò)細(xì)胞計(jì)數(shù)、流式凋亡檢測(cè)以及對(duì)凋亡發(fā)生的標(biāo)志性蛋白Cleaved Caspase-3蛋白水平的變化進(jìn)行檢測(cè),高表達(dá)SLC25A10之后明顯抑制肝癌細(xì)胞SMMC-7721凋亡的發(fā)生,并且在用經(jīng)典凋亡誘導(dǎo)藥物Etoposide處理之后,可以更明顯的抑制凋亡的發(fā)生。而在肝癌細(xì)胞HepG2中低表達(dá)SLC25A10之后進(jìn)行上述實(shí)驗(yàn),獲得了與高表達(dá)組正好相反的結(jié)果。在隨后的體內(nèi)實(shí)驗(yàn)中,裸鼠皮下成瘤同樣證實(shí)了高表達(dá)SLC25A10之后能夠抑制肝癌凋亡的發(fā)生。這其中的分子機(jī)制在于,高表達(dá)SLC25A10之后可以抑制線粒體途徑的凋亡發(fā)生,即減少線粒體內(nèi)cytochrome c的釋放,抑制cytochrome c和Apaf-1結(jié)合后與Caspase-9形成凋亡復(fù)合物,從而抑制下游的凋亡效應(yīng)蛋白如Caspase-3的激活,最終抑制凋亡的發(fā)生。綜上所述,在肝癌中高表達(dá)的SLC25A10是通過(guò)抑制線粒體凋亡通路參與到腫瘤的發(fā)生發(fā)展過(guò)程中的,SLC25A10可能作為治療肝癌的潛在靶點(diǎn),實(shí)現(xiàn)對(duì)肝癌的靶向治療。
[Abstract]:Worldwide, liver cancer as a common tumor, its morbidity and mortality are high. Data show that 50% of the world's new and death cases of liver cancer are from China. The World Health Organization estimates that without urgent action, about 10 million people in China will die from liver cancer between 2015 and 2030. At present, surgery is still the main method for the treatment of liver cancer, but the five-year survival rate of postoperative patients is only 15-40. Therefore, finding new treatment methods and new therapeutic targets is an urgent task in the clinical practice of liver cancer, and is also the research focus of this topic. SLC25A10, the tenth member of the A subfamily of Human Solute Carrier protein No. 25, is a transporter of mitochondrial membrane which transports dicarboxylate such as malic acid and succinic acid from the mitochondria to exchange phosphate. Sulfate and thiosulfate provide substrates for the processes of glycosylation and urea synthesis and thus maintain the distribution and homeostasis of intermediate products in and out of mitochondria during TCA cycle. In recent years, there have been many reports about the high expression of SLC25A10 in many kinds of tumors, but the expression level of SLC25A10 in HCC is rare, and its molecular mechanism is not clear. According to the previous experimental results, the transporter SLC25A10 is highly expressed in HCC and can inhibit the apoptosis of HCC. The purpose of this study is to further clarify the molecular mechanism of SLC25A10 inhibiting apoptosis in HCC. Find new potential targets for liver cancer and better target therapy for liver cancer. The expression of SLC25A10 was found to be high in clinical liver cancer tissues and paracancerous tissues by immunohistochemical analysis. At the same time, the results were further verified by Realtime-PCR and Western blot at mRNA and protein levels, and the same results were obtained. Then, the stable high expression of SLC25A10 in SMMC-7721 cells with lower SLC25A10 expression and the stable and low expression of SLC25A10 in HepG2 cells with higher SLC25A10 expression were detected by lentivirus infection. By cell count, flow cytometry and the change of Caspase-3 protein level, the expression of SLC25A10 significantly inhibited the apoptosis of SMMC-7721 cells, and the expression of SLC25A10 significantly inhibited the apoptosis of SMMC-7721 cells, and the expression of SLC25A10 significantly inhibited the apoptosis of SMMC-7721 cells. After treated with Etoposide, a classical apoptosis inducing drug, apoptosis was inhibited more obviously. The results were opposite to those in the high expression group after the low expression of SLC25A10 in HepG2 cells. In subsequent experiments in vivo, subcutaneous tumorigenesis in nude mice also confirmed that high expression of SLC25A10 could inhibit apoptosis of HCC. The molecular mechanism is that the overexpression of SLC25A10 can inhibit the apoptosis of mitochondrial pathway, that is, decrease the release of cytochrome c in mitochondria, inhibit the binding of cytochrome c and Apaf-1 to Caspase-9 and form apoptotic complex. Thus inhibiting the activation of downstream apoptosis-effector proteins such as Caspase-3 and ultimately inhibiting the occurrence of apoptosis. In conclusion, the high expression of SLC25A10 in HCC may be a potential target for the treatment of HCC by inhibiting the mitochondrial apoptotic pathway and participating in the process of tumor development.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 Piotr Stefaniuk;Janusz Cianciara;Alicja Wiercinska-Drapalo;;Present and future possibilities for early diagnosis of hepatocellular carcinoma[J];World Journal of Gastroenterology;2010年04期

，

本文編號(hào)：2096566