【摘要】：背景:SIRT家族(SIRT1-7)是一個(gè)NAD+-依賴(lài)的去乙�；�、乙酰化酶以及ADP-核糖基轉(zhuǎn)移酶家族,幾乎所有SIRT家族成員都被認(rèn)為在腫瘤形成中起到重要作用。最近有研究表明,SIRT4能通過(guò)調(diào)節(jié)谷氨酰胺代謝而具有抑癌基因的作用,但目前還沒(méi)有直接在人體組織蛋白水平進(jìn)行的SIRT4與人體腫瘤的相關(guān)性研究。方法:我們通過(guò)RT-PCR檢測(cè)了16對(duì)人結(jié)腸癌和癌旁正常結(jié)腸組織標(biāo)本中SIRT4的mRNA表達(dá)水平,然后進(jìn)一步分析了TCGA數(shù)據(jù)庫(kù)中包含223例樣本的人結(jié)腸癌表達(dá)譜芯片數(shù)據(jù)中SIRT4的mRNA表達(dá)水平,接下來(lái)分別對(duì)包含89例人結(jié)腸癌組織和75例人胃癌組織的組織芯片進(jìn)行免疫組化實(shí)驗(yàn)并分析SIRT4表達(dá)與人結(jié)腸癌和胃癌患者的臨床病理參數(shù)之間的關(guān)系。我們通過(guò)慢病毒構(gòu)建過(guò)表達(dá)SIRT4的人結(jié)腸癌細(xì)胞RKO和HT29的穩(wěn)定株,分析了過(guò)表達(dá)SIRT4對(duì)人結(jié)腸癌RKO和HT29的增殖、克隆形成的影響,并進(jìn)一步分析過(guò)表達(dá)SIRT4之后這些細(xì)胞對(duì)葡萄糖代謝抑制劑2-DG以及化療藥5-FU的敏感性的影響。我們還分析了過(guò)表達(dá)SIRT4對(duì)RKO和HT29細(xì)胞在谷氨酰胺缺乏培養(yǎng)基中存活率的影響。結(jié)果:SIRT4在結(jié)腸癌和胃癌組織中的表達(dá)水平顯著低于癌旁正常組織,SIRT4表達(dá)降低提示結(jié)腸癌和胃癌更差的病理分化程度(分別為:p=0.01,p=0.002),同時(shí)SIRT4表達(dá)低的結(jié)腸癌患者比SIRT4表達(dá)高的結(jié)腸癌患者術(shù)后5年生存率更差(p=0.0306)。過(guò)表達(dá)SIRT4抑制人結(jié)腸癌細(xì)胞RKO和HT29的增殖,并增加它們對(duì)葡萄糖代謝抑制劑2-DG以及化療藥5-FU的敏感性,同時(shí)過(guò)表達(dá)SIRT4減少了RKO和HT29細(xì)胞在谷氨酰胺缺乏培養(yǎng)基中的存活率。結(jié)論:SIRT4在人結(jié)腸癌和胃癌中扮演了抑癌基因的角色,SIRT4與人結(jié)腸癌和胃癌的病理分化程度相關(guān),與人結(jié)腸癌患者的術(shù)后5年生存率相關(guān)。SIRT4增加人結(jié)腸癌細(xì)胞對(duì)葡萄糖抑制劑2-DG和化療藥5-FU的敏感性。SIRT4對(duì)人結(jié)腸癌細(xì)胞的抑制作用不僅僅是通過(guò)抑制谷氨酰胺代謝引起的。SIRT4是人結(jié)腸癌和胃癌一個(gè)很有希望的的診斷標(biāo)志物,是人結(jié)腸癌一個(gè)很好的治療靶點(diǎn)。
[Abstract]:Background: SIRT family (SIRT1-7) is a NAD-dependent deacetylase, acetylase and ADP- ribonyltransferase family. Almost all members of SIRT family are considered to play an important role in tumor formation. Recent studies have shown that SIRT4 can inhibit oncogene by regulating glutamine metabolism, but there is no direct study on the relationship between SIRT4 and human tumor at human tissue protein level. Methods: we detected the mRNA expression of SIRT4 in 16 pairs of human colon cancer and adjacent normal colon tissues by RT-PCR, and further analyzed the mRNA expression level of SIRT4 in the TCGA database containing 223 samples of human colon cancer expression microarray. Then the tissue microarray containing 89 cases of human colon cancer and 75 cases of human gastric carcinoma were examined by immunohistochemistry and the relationship between the expression of SIRT4 and the clinicopathological parameters of patients with human colon cancer and gastric cancer was analyzed. We constructed human colon cancer cells RKO and HT29 stably by lentivirus and analyzed the effect of overexpression of SIRT4 on the proliferation and clone formation of RKO and HT29 in human colon cancer. The effects of overexpression of SIRT4 on the sensitivity of these cells to glucose metabolism inhibitor 2-DG and chemotherapeutic agent 5-FU were further analyzed. We also analyzed the effect of overexpression of SIRT4 on the survival rate of RKO and HT29 cells in glutamine deficient medium. Results: the expression level of SIRT4 in colon cancer and gastric cancer was significantly lower than that in adjacent normal tissues. The decrease of SIRT4 expression indicated the worse pathological differentiation of colon cancer and gastric cancer (p0. 01 p0. 002, respectively). Meanwhile, the expression of SIRT4 in colon cancer patients with low SIRT4 expression was higher than that in SIRT4 table. The 5-year survival rate of colon cancer patients was significantly lower than that of patients with colon cancer (p0. 0306). Overexpression of SIRT4 inhibited the proliferation of RKO and HT29 in human colon cancer cells and increased their sensitivity to glucose metabolism inhibitor 2-DG and chemotherapeutic drug 5-FU. Overexpression of SIRT4 reduced the survival rate of RKO and HT29 cells in glutamine deficient medium. Conclusion: SIRT4 plays an important role as a tumor suppressor gene in human colon cancer and gastric cancer. SIRT4 is related to the degree of pathological differentiation of human colon cancer and gastric cancer. SIRT4 increases the sensitivity of human colon cancer cells to glucose inhibitor 2-DG and chemotherapeutic agent 5-FU. The inhibitory effect of SIRT4 on human colon cancer cells is not only by inhibiting glutamine metabolism. SIRT4 is a promising diagnostic marker for colon and stomach cancer. It is a good therapeutic target for human colon cancer.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R735

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