本文選題：腺病毒 + 肝細(xì)胞癌��； 參考：《重慶醫(yī)科大學(xué)》2016年博士論文

【摘要】：目的:本研究探討共表達(dá)XAF-1和TFN-α對(duì)HCC細(xì)胞生長(zhǎng)的作用,并分析其分子機(jī)制。方法:在本研究中,檢測(cè)了肝細(xì)胞癌癌組織樣本中XAF-1的mRNA和蛋白水平,并在肝癌細(xì)胞系中證實(shí)了共表達(dá)XAF-1和TFN-α的腺病毒對(duì)肝癌細(xì)胞的顯著的抑制作用。建立HepG2 M人肝細(xì)胞癌97H細(xì)胞裸鼠移植瘤模型,并運(yùn)用重組共表達(dá)XAF-1和TNF-α腺病毒及Ad-con對(duì)裸鼠移植瘤模型進(jìn)行瘤內(nèi)注射,對(duì)照組每次注射同等體積的生理鹽水作為對(duì)照。觀察裸鼠移植瘤的生長(zhǎng)情況,并通過比較腫瘤體積的變化和對(duì)腫瘤的抑制率來評(píng)價(jià)其體內(nèi)治療的效果。結(jié)果:肝細(xì)胞癌癌組織樣本中XAF-1的表達(dá)量明顯低于肝細(xì)胞癌瘤旁樣本,說明其降低與腫瘤惡性程度相關(guān)。同時(shí)也在肝細(xì)胞癌細(xì)胞系,如MHCC 97L,HepG2和MHCC 97H中證實(shí)了XAF-1表達(dá)量的降低。本研究用一段2A肽編碼序列,構(gòu)建重組的腺病毒Ad-XAF-1TNF-α,有效的共表達(dá)了XAF-1和TFN-α。當(dāng)Ad-XAF-1和TNF-α轉(zhuǎn)染MHCC 97L細(xì)胞后,XAF-1和TFN-α的mRNA和蛋白水平明顯升高。此外,確認(rèn)了被Ad-XAF-1TNF-α轉(zhuǎn)染的MHCC 97L細(xì)胞,細(xì)胞增值率明顯低于單獨(dú)用Ad-XAF-1或Ad-TNF-α病毒感染的細(xì)胞。與PBS對(duì)照組注射的裸小鼠瘤體積相比較,,Ad-XAF-1TNF-α組及Ad-con組裸小鼠的瘤體積增長(zhǎng),差異有統(tǒng)計(jì)學(xué)意義(P0.05);Ad-XAF-1TNF-α治療組裸小鼠瘤體積增長(zhǎng)最為緩慢,與Ad-con組裸小鼠的瘤體積相比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。Ad-XAF-1TNF-α注射后的第7天,抑瘤率最高,達(dá)到65.87%,隨后抑瘤率呈下降趨勢(shì)。在注射后第28天,抑瘤率為60.78%.免疫組化染色以及RT-PCR實(shí)驗(yàn)結(jié)果表明,與Ad-con組及PBS組裸鼠瘤組織XAF-1、TNF-α的陽性表達(dá)率相比(6.11%±1.73,6.42%±2.59),Ad-XAF-1TNF-α治療組裸鼠瘤組織XAF-1、TNF-α的陽性率表達(dá)增高(16.39%±1.39),差異有統(tǒng)計(jì)學(xué)意義(P0.05)。Ad-XAF-1TNF-α治療組裸鼠瘤組織中XAF-1、TNF-α基因的mRNA表達(dá)水平均較Ad-con組、PBS組裸鼠瘤組織中XAF-1、TNF-α基因的mRNA表達(dá)水平增高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:Ad-XAF-1TNF-α共表達(dá)病毒可以較單表達(dá)XAF-1、TNF-α的腺病毒更有效的抑制肝細(xì)胞癌細(xì)胞增殖,介導(dǎo)XAF-1、TNF-α可在裸小鼠移植瘤模型內(nèi)進(jìn)行有效的表達(dá),減小瘤體積從而發(fā)揮抑制移植瘤生長(zhǎng)的作用,同時(shí)還能促進(jìn)XAF-1、TNF-α在移植瘤中的表達(dá)及mRNA水平的表達(dá),其有望成為一種有效抵抗肝細(xì)胞癌的抗腫瘤因子。
[Abstract]:Aim: to investigate the effect of coexpression of XAF-1 and TFN- 偽 on the growth of HCC cells and to analyze its molecular mechanism. Methods: in this study, the levels of mRNA and protein of XAF-1 in hepatocellular carcinoma (HCC) tissues were detected, and the inhibitory effect of adenovirus co-expressing XAF-1 and TFN- 偽 on HCC cells was confirmed. The transplanted tumor model of HepG2 M human hepatocellular carcinoma 97H cell in nude mice was established. The nude mice transplanted tumor model was injected with recombinant co-expression of XAF-1, TNF- 偽 adenovirus and Ad-con. The control group was injected with normal saline of the same volume each time. The growth of xenografts in nude mice was observed and the effect of treatment in vivo was evaluated by comparing the changes of tumor volume and tumor inhibition rate. Results: the expression of XAF-1 in hepatocellular carcinoma tissues was significantly lower than that in the adjacent samples, indicating that the decrease of XAF-1 expression was related to the malignancy of the tumor. The decrease of XAF-1 expression was also confirmed in hepatocellular carcinoma cell lines such as MHCC 97L HepG2 and MHCC 97H. In this study, the recombinant adenovirus Ad-XAF-1TNF- 偽 was constructed with a fragment of 2A peptide encoding sequence. The recombinant adenovirus Ad-XAF-1TNF- 偽 was effectively coexpressed with XAF-1 and TFN- 偽. When Ad-XAF-1 and TNF- 偽 were transfected into MHCC 97L cells, the mRNA and protein levels of XAF-1 and TFN- 偽 increased significantly. In addition, the proliferation rate of MHCC 97L cells transfected with Ad-XAF-1TNF- 偽 was significantly lower than that of cells infected with Ad-XAF-1 or Ad-TNF- 偽 alone. Compared with the nude mice injected with PBS, the tumor volume of Ad-XAF-1TNF- 偽 group and Ad-con group increased significantly (P 0.05). The tumor volume of nude mice treated with Ad-XAF-1TNF- 偽 was the slowest, and the tumor volume of Ad-con group was higher than that of Ad-con group. On the 7th day after injection, the tumor inhibition rate was the highest, reaching 65.87, and then decreased. On the 28th day after injection, the tumor inhibition rate was 60.78%. Immunohistochemical staining and RT-PCR assay showed that, Compared with Ad-con group and PBS group, the positive rate of XAF-1TNF- 偽 in tumor tissue of nude mice was increased by 6.11% 鹵1.73 鹵6.42% 鹵2.59 ~ XAF-1TNF- 偽 treatment group. The positive rate of XAF-1TNF- 偽 expression in tumor tissue of nude mice in Ad-con group and PBS group was 16.39% 鹵1.39% higher than that in Ad-con group. There was significant difference in the expression of XAF-1TNF- 偽 gene in tumor tissue of nude mice treated with P0.05. Ad-XAF-1TNF- 偽. The mRNA expression level of XAF-1TNF- 偽 gene was higher than that of Ad-con group. The mRNA expression level of XAF-1TNF- 偽 gene was increased in nude mice in PBS group. The difference was statistically significant (P 0.05). Conclusion compared with the adenovirus expressing XAF-1TNF- 偽 alone, the adenovirus can inhibit the proliferation of hepatocellular carcinoma more effectively, and mediate the expression of XAF-1TNF- 偽 in nude mice transplanted tumor model, and reduce the volume of tumor and thus play a role in inhibiting the growth of transplanted tumor. It can also promote the expression of XAF-1TNF- 偽 and the expression of mRNA in transplanted tumors, which is expected to be an effective anti-tumor factor against hepatocellular carcinoma.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R735.7

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本文編號(hào)：1840976