本文選題：卟啉 + 光動(dòng)力療法　； 參考：《遵義醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的：觀察新合成的金屬卟啉化合物對(duì)肝癌細(xì)胞(HepG2)、胃癌細(xì)胞(SGC-7901)、結(jié)腸癌細(xì)胞(sw480)、高分化鼻咽癌細(xì)胞(CNE-1)、低分化鼻咽癌細(xì)胞(CNE-2)、小鼠單核巨噬細(xì)胞(RAW264.7)、肺癌細(xì)胞(A549)以及乳腺癌細(xì)胞(MCF-7)的抗腫瘤作用，并篩選療效較好的藥物進(jìn)一步研究卟啉化合物抗腫瘤作用機(jī)理。 方法：1.藥物抗腫瘤活性篩選：SRB法和MTT法測(cè)定藥物對(duì)腫瘤細(xì)胞增殖的影響；鄰二氮菲-Fe2+氧化法和鄰苯三酚自氧化法測(cè)定藥物對(duì)腫瘤細(xì)胞清除羥自由基和超氧陰離子自由基能力的影響；Griess法測(cè)定腫瘤細(xì)胞中NO自由基含量的變化；Giemsa染色法觀察腫瘤細(xì)胞形態(tài)學(xué)的變化。2.藥物抗腫瘤機(jī)理研究：羅丹明123測(cè)定線粒體膜電位變化；流式細(xì)胞儀檢測(cè)細(xì)胞凋亡；DNA完整性檢測(cè)；實(shí)時(shí)熒光定量PCR法檢測(cè)腫瘤細(xì)胞C-myc基因的mRNA表達(dá)量的變化；ELISA法測(cè)定腫瘤細(xì)胞端粒酶活性的變化。 結(jié)果：八種金屬卟啉化合物對(duì)腫瘤細(xì)胞的生長(zhǎng)均具有一定的抑制作用，且隨著藥物濃度的升高，，細(xì)胞的生長(zhǎng)抑制率基本呈上升的趨勢(shì)；細(xì)胞對(duì)羥自由基和超氧陰離子自由基的清除能力下降；細(xì)胞上清液中NO自由基的含量升高；Giemsa染色后觀察到凋亡細(xì)胞數(shù)目明顯增加且藥物對(duì)HepG2細(xì)胞作用最顯著。上述效果最好的兩種藥物是Rup-03和Rup-04，Rup-03對(duì)五種腫瘤細(xì)胞(HepG2、CNE-1、CNE-2、sw480以及RAW264.7細(xì)胞)的殺傷作用較好，Rup-04對(duì)SGC-7901細(xì)胞的殺傷作用較好。隨著Rup-03濃度的增加(10-9～10-6mol/L)，線粒體膜電位熒光強(qiáng)度減弱；流式細(xì)胞儀顯示HepG2細(xì)胞凋亡率逐漸增加；瓊脂糖凝膠電泳圖顯示DNA發(fā)生了降解；C-myc基因的mRNA相對(duì)表達(dá)量減少且端粒酶活性降低。這些結(jié)果均表明，細(xì)胞凋亡程度隨著藥物濃度的增加而加重。 結(jié)論：八種新型金屬卟啉化合物對(duì)腫瘤細(xì)胞均具有一定的殺傷作用，且殺傷作用與濃度具有一定相關(guān)性，總體效果最好的藥物為Rup-03，適宜濃度為10-9～10-6mol/L。該新型卟啉化合物的抗腫瘤作用機(jī)理可能是經(jīng)光照后生成自由基，進(jìn)而抑制腫瘤細(xì)胞C-myc基因的表達(dá)以及端粒酶活性并誘導(dǎo)細(xì)胞發(fā)生凋亡。
[Abstract]:Objective: to observe the effects of newly synthesized metalloporphyrin compounds on hepatocellular carcinoma cell line HepG2, gastric cancer cell line SGC-7901, colon cancer cell line SW480, well-differentiated nasopharyngeal carcinoma cell line CNE-1, poorly differentiated nasopharyngeal carcinoma cell line CNE-2, mouse mononuclear macrophage RAW264.7, lung cancer cell A549) and breast cancer. The antitumor effect of MCF-7), The mechanism of anti-tumor action of porphyrin compounds was further studied. Method 1: 1. The effects of antitumor drugs on the proliferation of tumor cells were determined by the method of antitumor activity: the effects of phenanthrene Fe2 oxidation and pyrogallol autooxidation on the ability of scavenging hydroxyl radical and superoxide anion free radical in tumor cells were measured. The changes of no Free radical content in tumor cells by Griess method; the morphological changes of tumor cells were observed by Giemsa staining. The mechanism of drug anti-tumor: Rhodamine 123 was used to detect mitochondrial membrane potential, flow cytometry was used to detect apoptosis and DNA integrity, real-time fluorescence quantitative PCR was used to detect the mRNA expression of C-myc gene in tumor cells. Telomerase activity of tumor cells was determined by Elisa. Results: all of the eight metalloporphyrin compounds could inhibit the growth of tumor cells, and the growth inhibition rate of tumor cells increased with the increase of drug concentration. The activity of scavenging hydroxyl radical and superoxide anion radical was decreased, and the content of no free radical in supernatant increased after Giemsa staining, and the number of apoptotic cells was obviously increased, and the effect of drugs on HepG2 cells was most significant. The two most effective drugs were Rup-03 and Rup-04 Rup-03. Rup-04 had better killing effect on SGC-7901 cells than Rup-04 on five kinds of tumor cells (HepG2CNE-1 CNE-2Osw480 and RAW264.7). With the increase of Rup-03 concentration, the fluorescence intensity of mitochondrial membrane potential decreased, the apoptosis rate of HepG2 cells increased by flow cytometry, the DNA degradation occurred by agarose gel electrophoresis. The relative expression of C myc mRNA and telomerase activity were decreased. These results indicate that the degree of apoptosis increases with the increase of drug concentration. Conclusion: all the eight new metalloporphyrin compounds have a certain killing effect on tumor cells, and the killing effect is related to the concentration of Rup-03. the best drug is Rup-03, and the optimum concentration is 10-9 ~ 10-6 mol 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1). The anti-tumor mechanism of the new porphyrin compound may be the formation of free radicals after irradiation, which can inhibit the expression of C-myc gene, telomerase activity and induce apoptosis of tumor cells.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R96

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