本文選題：CRC + BRC��； 參考：《昆明醫(yī)科大學》2017年博士論文

【摘要】：第一部分TNF-α-308 G/A多態(tài)性與結直腸癌和乳腺癌患病風險的研究[目的]TNFα是一個炎癥相關的細胞因子,它參與多種癌癥的發(fā)生發(fā)展過程。目前TNF-α-308G/A的多態(tài)性與結直腸癌(CRC)和乳腺癌(BRC)的相關性尚不完全清楚,尤其是與CRC、BRC的臨床特征、實驗室指標的關聯(lián)性尚未有深入的研究。本實驗中,綜合CRC和BRC各自的特點,把疾病分為不同的亞型、將傳統(tǒng)組織病理學分型與基因分型相結合,研究TNFα基因多態(tài)性與CRC和BRC發(fā)生與轉移的風險從而指導個體化的診斷與治療。[方法]研究收錄昆明醫(yī)科大學第一附屬醫(yī)院和云南省腫瘤醫(yī)院570例CRC、490例BRC患者及相應的無病對照。收集相應的人口學資料和臨床資料;對照組收集人口學資料。采用高通量時間飛行質譜生物芯片系統(tǒng)檢測實驗對象TNFα的單核苷酸多態(tài)性(SNP)。通過比較病例組與對照組人口學特征、CRC和BRC各亞型間的臨床特征和實驗室指標及各等位基因分布頻率的差異,計算OR及95%CI,從而比較不同基因型與CRC和BRC發(fā)生及轉移的相對危險度。[結果]TNFα基因rs1800629位點檢測到GG、AA及GA三種基因型。結直腸癌中,年齡和性別在病例組和對照組中滿足頻率匹配(P=0.460.77),TNF-α-308 G和A的等位基因頻率符合哈迪溫伯格平衡定律(PHWE=0.850.67)。BRC中病例組和對照組之間的年齡分布無顯著差異(P=0.065),G和A的等位基因頻率符合哈迪溫伯格平衡定律(PHWE=0.870.96)。在所有的CRC患者中,TNF-α-308 GA 增加了 CRC 患者遠處轉移的風險(OR=2.911,95%CI:1.697-4.992,P= 0.00017),經(jīng)多因素Logistic回歸分析,該數(shù)值仍具有顯著的差異(OR=2.947,95%CI:1.515-5.572,P=0.00121)。有趣的是,將CRC分組后,我們發(fā)現(xiàn)GA和AA僅僅是直腸癌遠處轉移的危險因素(OR=4.481,95%CI:2.072-9.693,P=0.00025)而和結腸癌的遠處轉移發(fā)生沒有確切相關性(OR=1.685,95%CI:0.592-4.796),校正所有臨床資料后該數(shù)值仍具有顯著的差異(OR =7.099,95%CI:2.482-20.301,P= 0.000256)。GA和AA基因型是CRC患者發(fā)生肥胖、腫瘤體積增大的危險因素,攜帶GA和AA基因型的患者的肥胖風險較攜帶GG基因型增加(OR = 2.056,95%CI:1.197-3.530,P=0.01);攜帶 GA 和 AA 型的 CRC 具有更大的腫瘤體積(OR=2.225,95%CI:1.049-4.719,P=0.044)。TNF-α-308 GA是CRC患者中性粒細胞增高的危險因素(OR= 4.764,95%CI:2.39-9.495,P=0.000012)。BRC患者中,GA和AA基因型是腫瘤體積的保護因素,攜帶GA和AA基因型的BRC患者腫瘤體積相對較小(OR = 0.137,95%CI:0.019-1.013,P=0.023),經(jīng)多因素Logistic回歸分析后,結果仍然有意義(OR = 0.115,95%CI:0.013-0.991,P=0.023)。在TNBC中,GA和AA基因型更頻繁的出現(xiàn)于初潮年齡大于15歲的患者(OR =11.75,95%CI:1.761-78.416,P=0.003)及罹患高血壓的患者中(OR=8,95%CI:1.02-62.737,P=0.025)。在 Luminal-A 型患者中,右側乳腺癌的GA和AA基因型頻率較高(OR=5.073,95%CI:1.022-25.194,P=0.031),HER-2患者中,具有GA和AA基因型的患者初次診斷乳腺癌的年齡偏大(OR= 1.211,95%CI:1.004-1.46,P=0.029)。通過比較實驗室指標基因型的分布差異,揭示了 GA和AA基因型與白蛋白和膽固醇水平的相關性,TNF-α-308 GA 是 BRC 低蛋白血癥(OR= 2.571,95%CI:1.194-5.535,P=0.013)及 TNBC高膽固醇血癥(OR= 7.75,95%CI:0.842-69.708,P=0.041)的危險因素。[結論]TNF-α-308 GA多態(tài)性是CRC發(fā)生遠處轉移的危險因素,尤其增加了直腸癌患者發(fā)生遠處轉移的風險,而和乳腺癌的復發(fā)轉移沒有明顯相關性,GA和AA基因型是CRC患者發(fā)生肥胖、腫瘤體積增大的危險因素,提示TNFα基因rs 1800629位點是影響CRC預后的重要因素之一。GA和AA基因型增加了BRC低蛋白血癥和TNBC高膽固醇血癥、TNBC罹患高血壓及Luminal-A患者右乳癌的患病風險,具有GA和AA基因型的患者有較小的腫瘤體積,提示TNF-α-308 GA的多態(tài)可以預測腫瘤的原發(fā)部位、大小及評估BRC的營養(yǎng)狀況。第二部分Dexamethasone誘導三陰性乳腺癌耐藥的機制研究[目的]研究地塞米松(Dex)在促進三陰性乳腺癌(TNBC)耐藥中的機制[方法]用SRB實驗檢測Dex對TNBC細胞經(jīng)化療藥物順鉑(Cpt)、多西他賽(DTX)處理后對生長、增殖的影響;用免疫印跡實驗檢測糖皮質激素(GCs)對KLF5蛋白的誘導,以及相關凋亡蛋白的檢測;用real-time PCR、雙熒光素酶報告基因實驗檢測地塞米松對KLF5的轉錄活性的影響;用ChIP實驗驗證糖皮質激素受體(GR)對KLF5啟動子調控的具體位點;用流式細胞儀檢測Dex通過KLF5介導的TNBC細胞凋亡的影響;用裸鼠成瘤實驗在體內(nèi)證明Dex在TNBC對DTX敏感性中的影響。[結果]Dex可以促進TNBC細胞HCC1937和HCC1806的生長和增殖,同時我們發(fā)現(xiàn)其誘導KLF5在乳腺癌細胞系中的表達;在TNBC細胞中,Dex可以促進KLF5的轉錄活性,活化GR并促其結合至KLF5啟動子區(qū)域的Sp1位點,增強KLF5的啟動子的活性,進而促進KLF5的轉錄表達;敲低KLF5能夠抑制三陰性乳腺癌細胞的存活;KLF5介導了 Dex對DDTX和Cpt引起的細胞凋亡的抑制作用;Dex降低了乳腺腫瘤在動物體內(nèi)對化療藥物的敏感性,而沉默KLF5表達則回復了腫瘤對DTX的敏感性;這些結果說明Dex至少部分通過KLF5調控三陰性乳腺癌細胞對DTX和Cpt的耐藥。[結論]我們證實了 Dex通過GR調控下游靶基因KLF5的轉錄,進而促進TNBC細胞的增殖;Dex通過KLF5的表達,進而促進了 TNBC對化療藥物的耐藥。說明,KLF5是三陰性乳腺癌治療和預測療效潛在的靶點和生物標記物。
[Abstract]:Part one TNF- alpha -308 G/A polymorphism and the risk of colorectal and breast cancer [Objective]TNF alpha is an inflammatory cytokine that is involved in the development of a variety of cancers. The association of TNF- alpha -308G/A with colorectal cancer (CRC) and breast cancer (BRC) is not completely clear, especially with CRC, BRC. The correlation of clinical features and laboratory indicators has not been studied in depth. In this experiment, the characteristics of CRC and BRC were divided into different subtypes, and the traditional histopathological and genotyping were combined to study the risk of TNF alpha gene polymorphism and the occurrence and transfer of CRC and BRC and to guide the individualized diagnosis and treatment. Methods: the study included 570 cases of CRC, 490 Cases of BRC patients and the corresponding disease-free control in the First Affiliated Hospital of Kunming Medical University and Yunnan cancer hospital. The corresponding demographic data and clinical data were collected. The control group collected human data. The high throughput time flight mass spectrometry biochip system was used to detect the single nucleotide polymorphisms of TNF alpha. SNP. By comparing the demographic characteristics of the case group with the control group, the differences in the clinical features, the laboratory indexes and the frequency of the distribution of the alleles in the CRC and BRC subtypes, the OR and 95%CI were calculated, and the relative risk of the occurrence and metastasis of CRC and BRC was compared. [results] the rs1800629 loci of]TNF a gene detected GG, AA and GA. Three genotypes. In colorectal cancer, age and sex meet frequency matching (P=0.460.77) in case group and control group. The allele frequency of TNF- alpha -308 G and A is consistent with Hardy Weinberg equilibrium law (PHWE=0.850.67).BRC between case group and control group, there is no significant difference in age distribution (P=0.065), G and A allele frequency conforms to ha. Di Weinberg's law of equilibrium (PHWE=0.870.96). In all CRC patients, TNF- alpha -308 GA increases the risk of distant metastasis of CRC patients (OR=2.911,95%CI:1.697-4.992, P= 0.00017). The value of this value is still significant (OR=2.947,95% CI:1.515-5.572, P=0.00121). We found that GA and AA were only a risk factor for distant metastasis of rectal cancer (OR=4.481,95%CI:2.072-9.693, P=0.00025) and had no definite correlation with distant metastasis of colon cancer (OR=1.685,95%CI:0.592-4.796). After correcting all clinical data, the value still had significant difference (OR =7.099,95%CI:2.482-20.301, P= 0.000256).GA and AA base. The risk factors for obesity and tumor volume increased in CRC patients. The risk of obesity carrying GA and AA genotype was higher than that of GG genotype (OR = 2.056,95%CI:1.197-3.530, P=0.01), and CRC with GA and AA type CRC had greater tumor volume (OR=2.225,95%CI: 1.049-4.719,) In OR= 4.764,95%CI:2.39-9.495 (P=0.000012).BRC patients, the GA and AA genotypes are the protective factors of tumor volume. The volume of the tumor carrying GA and AA genotype is relatively small (OR = 0.137,95%CI:0.019-1.013, P=0.023). CI:0.013-0.991, P=0.023). In TNBC, the GA and AA genotypes appear more frequently in patients aged more than 15 years old (OR =11.75,95%CI:1.761-78.416, P=0.003) and patients with hypertension (OR=8,95%CI:1.02-62.737, P=0.025). In Luminal-A type patients, the frequency of GA and genotypes in right side breast cancer is higher. -25.194, P=0.031), in patients with HER-2, the age of the first diagnosis of breast cancer with GA and AA genotypes was larger (OR= 1.211,95%CI:1.004-1.46, P=0.029). The correlation between the genotype of GA and AA was revealed by comparing the distribution differences in the laboratory index genotypes. 2.571,95%CI:1.194-5.535, P=0.013) and the risk factors of TNBC hypercholesterolemia (OR= 7.75,95%CI:0.842-69.708, P=0.041). [conclusion]TNF- alpha -308 GA polymorphism is a risk factor for distant metastasis of CRC, especially increasing the risk of distant metastasis in rectal cancer patients, but not associated with recurrence and metastasis of breast cancer, GA and AA. Genotype is a risk factor for obesity and tumor volume in CRC patients. It is suggested that the RS 1800629 locus of TNF a gene is one of the important factors affecting the prognosis of CRC. The.GA and AA genotypes increase BRC hypoproteinemia and TNBC hypercholesterolemia, and the risk of high blood pressure and Luminal-A patients with right breast cancer with GA and AA genotypes The patient has a small tumor volume, suggesting that TNF- alpha -308 GA polymorphism can predict the primary site of the tumor, size and evaluation of the nutritional status of BRC. Second the mechanism of Dexamethasone induced resistance to three negative breast cancer [Objective] to study the mechanism of dexamethasone (Dex) in promoting the resistance of three negative breast cancer (TNBC) [method] with SRB test The effects of Dex on the growth and proliferation of TNBC cells treated with chemotherapeutic drugs cisplatin (Cpt) and docetaxel (DTX) were detected. The induction of KLF5 protein by Glucocorticoid (GCs) and the detection of related apoptotic proteins were detected by immunoblot test, and the transcriptional activity of dexamethasone was detected by real-time PCR and double fluoroenzyme reporter gene test. Influence; the specific site of glucocorticoid receptor (GR) Regulation of KLF5 promoter was verified by ChIP test; the effect of Dex on the apoptosis of TNBC cells mediated by KLF5 was detected by flow cytometry; the effect of Dex in TNBC on DTX sensitivity was demonstrated in nude mice in vivo. [results] Dex could promote the growth of TNBC cell HCC1937 and the growth of TNBC. Proliferation, and we found that it induces the expression of KLF5 in the breast cancer cell lines; in TNBC cells, Dex can promote the transcriptional activity of KLF5, activate GR and bind to the Sp1 loci of the KLF5 promoter region, enhance the activity of the promoter of KLF5, and promote the transcriptional table of KLF5; knockdown KLF5 can inhibit the survival of three negative breast cancer cells. Live; KLF5 mediates the inhibitory effect of Dex on cell apoptosis induced by DDTX and Cpt; Dex reduces the sensitivity of breast tumors to chemotherapeutic drugs in animals, while silent KLF5 expression revert to the tumor's sensitivity to DTX; these results indicate that Dex at least partly controls the resistance of three negative breast cancer cells to DTX and Cpt. [Conclusion] We confirm that Dex regulates the transcription of the downstream target gene KLF5 by GR and thus promotes the proliferation of TNBC cells. Dex promotes the resistance of TNBC to chemotherapeutic drugs through KLF5 expression, indicating that KLF5 is a potential target and biomarker for the treatment and prediction of three negative breast cancer.

【學位授予單位】：昆明醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R737.9;R735.34

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相關期刊論文 前2條

1 Xu-Feng Guo;Jun Wang;Shi-Jie Yu;Jia Song;Meng-Yao Ji;Zhuo Cao;Ji-Xiang Zhang;Jing Wang;Wei-Guo Dong;;TNF-a-308 polymorphism and risk of digestive system cancers:A meta-analysis[J];World Journal of Gastroenterology;2013年48期

2 鄭瑩;吳春曉;張敏璐;;乳腺癌在中國的流行狀況和疾病特征[J];中國癌癥雜志;2013年08期

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本文編號：1809669