本文選題：肝癌細(xì)胞 + 自體輸血　； 參考：《西南醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:肝細(xì)胞肝癌在腫瘤導(dǎo)致死亡的疾病中排名世界第三,特別是在發(fā)展中國(guó)家,發(fā)病率還呈上升的趨勢(shì)。手術(shù)治療是根治肝癌的最有效的手段,但是僅有10%-30%的患者在診斷出肝癌時(shí)有手術(shù)指征。肝癌患者進(jìn)行肝切除術(shù)或者肝移植手術(shù)時(shí)常常會(huì)伴隨著大量失血,往往需要輸血,異體輸血存在著很多弊端,比如不良輸血反應(yīng),免疫抑制以及疾病傳播等,而特別是對(duì)于腫瘤患者來(lái)講,可能還會(huì)引起腫瘤的復(fù)發(fā),因此推薦肝癌患者進(jìn)行自體輸血。但是肝癌患者自體輸血一直以來(lái)又存在著爭(zhēng)議,有學(xué)者擔(dān)心血中可能存在的肝癌細(xì)胞有可能會(huì)引起腫瘤的血行轉(zhuǎn)移。故本實(shí)驗(yàn)通過(guò)研究γ射線對(duì)人肝癌細(xì)胞的增殖、遷移、侵襲、凋亡的影響,來(lái)探究γ射線對(duì)肝癌患者的自體血液中殘留的腫瘤細(xì)胞的滅活效果,尋找合適的輻照劑量,為肝癌患者的自體輸血的安全應(yīng)用提供依據(jù)。方法:1)人肝癌細(xì)胞株SMMC7721和HepG2的體外培養(yǎng)。2)用不同劑量的γ射線處理HCC(Hepatoma carcinoma cell)后,用CCK8(cell counting kit-8)實(shí)驗(yàn)檢測(cè)細(xì)胞的增殖能力,吸光度值的大小可以反應(yīng)活細(xì)胞的相對(duì)數(shù)量;劃痕實(shí)驗(yàn)檢測(cè)細(xì)胞的遷移能力,細(xì)胞移動(dòng)的距離可以代表遷移能力;Transwell實(shí)驗(yàn)以及MPP-9蛋白表達(dá)量檢測(cè)來(lái)反應(yīng)細(xì)胞的侵襲能力,穿過(guò)的細(xì)胞越多,MMP-9蛋白的表達(dá)越多,細(xì)胞的侵襲能力越強(qiáng);annexin V/PI染色(流式細(xì)胞術(shù))和P53凋亡蛋白的表達(dá)量測(cè)定來(lái)檢測(cè)細(xì)胞的凋亡情況。3)用SPSS17.0軟件統(tǒng)計(jì)分析實(shí)驗(yàn)結(jié)果,P0.05表示差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:1)γ射線處理細(xì)胞后,細(xì)胞失去了正常形態(tài),出現(xiàn)了細(xì)胞變形,漂浮,細(xì)胞膜不完整,胞質(zhì)內(nèi)有空泡,細(xì)胞碎裂等一系列變化;2)γ射線處理后24h,48h,72h,細(xì)胞的增殖能力都明顯受到抑制,細(xì)胞活力跟培養(yǎng)時(shí)間和γ射線劑量成反比,隨著培養(yǎng)時(shí)間和劑量增加,細(xì)胞活力降低,差異有統(tǒng)計(jì)學(xué)意義(P0.01);3)γ射線對(duì)細(xì)胞的遷移能力有抑制作用,24h劃痕實(shí)驗(yàn)結(jié)果顯示,隨著劑量增加,細(xì)胞遷移的距離減小,差異有統(tǒng)計(jì)學(xué)意義(P0.01);4)γ射線對(duì)細(xì)胞的侵襲能力有抑制作用,隨著劑量增加,穿過(guò)小室的細(xì)胞數(shù)量逐漸減小,差異具有統(tǒng)計(jì)學(xué)意義(P0.01);Western Blot法結(jié)果顯示,輻照后,MPP-9蛋白的表達(dá)量降低(P0.01);5)γ射線會(huì)抑制細(xì)胞的克隆形成,20Gy開始,細(xì)胞沒有克隆形成。6)流式凋亡檢測(cè)顯示,細(xì)胞早期凋亡率隨著γ射線的劑量增加而增加(P0.01),Western Blot法結(jié)果顯示,γ射線處理HCC后,P53凋亡蛋白的表達(dá)量增加(P0.01)。結(jié)論:γ射線可以有效地抑制肝癌細(xì)胞的增殖,遷移和侵襲,促進(jìn)細(xì)胞凋亡,可以有效地滅活肝癌細(xì)胞;推薦使用40Gy的劑量應(yīng)用于肝癌患者自體血液中腫瘤細(xì)胞的滅活,提高肝癌患者自體輸血的安全性和有效性。
[Abstract]:Objective: hepatocellular carcinoma (HCC) is the third leading cause of cancer death in the world, especially in developing countries.Surgical treatment is the most effective method to cure liver cancer, but only 10 to 30% of the patients have surgical indications in the diagnosis of liver cancer.Liver cancer patients undergoing hepatectomy or liver transplantation are often accompanied by massive blood loss and often need blood transfusions. Allogeneic blood transfusion has many drawbacks, such as adverse transfusion reactions, immunosuppression and disease transmission.Especially for cancer patients, it may also lead to recurrence of tumor, so we recommend autologous blood transfusion for liver cancer patients.However, autologous blood transfusion in patients with liver cancer has been controversial, some scholars worry that the existence of liver cancer cells in the blood may cause tumor blood metastasis.Therefore, the effect of 緯-ray on the proliferation, migration, invasion and apoptosis of human hepatoma cells was studied in order to explore the inactivation effect of 緯-ray on the residual tumor cells in autologous blood of patients with liver cancer, and to find the appropriate irradiation dose.To provide the basis for the safe application of autologous blood transfusion in patients with liver cancer.Methods in vitro culture of human hepatoma cell lines SMMC7721 and HepG2. (2) HCC(Hepatoma carcinoma cells were treated with different doses of 緯 -rays. The proliferative ability of HCC(Hepatoma carcinoma cells was measured by CCK8(cell counting kit-8). The relative number of living cells could be reflected by the size of absorbance.The migration ability of cells was measured by scratch test. The distance of cell movement could represent the migration ability and the expression of MPP-9 protein to reflect the invasion ability of cells. The more cells passed through, the more MMP-9 protein was expressed.The stronger the invasiveness of cells was, the more annexin V/PI staining (flow cytometry) and p53 apoptotic protein expression were measured to detect the apoptosis of cells.Results after the cell was treated with 緯 -ray, the cell lost its normal shape, appeared cell deformation, floating, incomplete cell membrane, and vacuole in the cytoplasm.A series of changes, such as cell fragmentation, etc.) after 24 h ~ 48 h ~ 72 h of 緯 -ray treatment, the cell proliferation ability was obviously inhibited, and the cell viability was inversely proportional to the culture time and 緯 -ray dose, and decreased with the increase of culture time and dose.The difference was statistically significant (P 0.01) 緯 -ray could inhibit the migration ability of cells. The results of 24 h scratch test showed that the distance of cell migration decreased with the increase of dose, and the difference was statistically significant (P 0.01) 緯 -ray could inhibit the invasion ability of cells.With the increase of dose, the number of cells passing through the cell gradually decreased. The results of Western Blot analysis showed that the expression of MPP-9 protein decreased after irradiation.Flow cytometry showed that the early apoptosis rate increased with the increase of 緯 -ray dose. The results of Western Blot showed that the expression of p53 apoptosis protein increased after 緯 -ray treatment of HCC.Conclusion: 緯-ray can effectively inhibit the proliferation, migration and invasion of hepatoma cells, promote cell apoptosis, and effectively inactivate hepatoma cells. It is recommended that the dose of 40Gy be used in the inactivation of tumor cells in autologous blood of patients with liver cancer.To improve the safety and efficacy of autologous blood transfusion in patients with liver cancer.
【學(xué)位授予單位】：西南醫(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

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本文編號(hào)：1731132