本文選題：口腔鱗狀細(xì)胞癌 + 雷公藤屬��； 參考：《華北理工大學(xué)》2017年碩士論文

【摘要】：目的通過研究雷公藤甲素(triptolide,TP)對人口腔鱗癌細(xì)胞株的增殖和10號染色體同源丟失性磷酸酶張力蛋白(phosphataseand tensin homolog,PTEN)基因表達(dá)的影響,探討TP抗口腔鱗狀癌的作用機(jī)制,以期為口腔鱗狀細(xì)胞癌的治療提供實(shí)驗(yàn)依據(jù)。方法以口腔鱗狀癌細(xì)胞作為研究對象,將口腔鱗狀癌細(xì)胞分為對照組(A)和四個(gè)實(shí)驗(yàn)組(B、C、D、E),分別給予濃度為0、1.25、2.5、5.0和10μg/L的TP進(jìn)行處理。1通過甲基噻唑基四唑(methyl thiazolyl tetrazolium method,MTT)染色法檢測24、48和72h后TP對口腔鱗狀癌細(xì)胞增殖變化的影響;2流式細(xì)胞檢測技術(shù)檢測48h后TP對口腔鱗狀癌細(xì)胞周期的影響;3實(shí)時(shí)熒光定量(Real-time Quantitative polymerase chain reaction,RT-PCR)檢測24、48和72h后TP對PTEN的信使核糖核酸(Messenger RNA m RNA)表達(dá)的影響;4蛋白印跡法(Western-blot)檢測72h后TP對PTEN蛋白表達(dá)的影響。結(jié)果1 MTT結(jié)果:24h后B、C、D、E組的細(xì)胞抑制率分別為(12.48±0.86)%、(23.61±0.10)%、(35.37±0.10)%、(46.58±0.91)%,與對照組相比,差異具有統(tǒng)計(jì)學(xué)意義(P0.05);72h后四個(gè)實(shí)驗(yàn)組的細(xì)胞抑制率分別為(26.92±0.14)%、(38.67±0.11)%、(72.62±0.89)%、(90.42±0.28)%,與對照組相比,差異有統(tǒng)計(jì)學(xué)意義(P0.05);48h后,細(xì)胞抑制率隨濃度增加而增加,統(tǒng)計(jì)分析結(jié)果顯示各組間兩兩比較均有統(tǒng)計(jì)學(xué)意義(P0.05);說明TP可抑制口腔鱗狀癌細(xì)胞增殖。2.流式細(xì)胞檢測技術(shù)結(jié)果顯示,TP作用口腔鱗狀癌細(xì)胞48后,A、B、C、D、E各組G1期細(xì)胞比例分別為(58.78±0.98)%、(70.85±0.41)%、(74.90±0.28)%、(79.14±0.45)%、(84.13±0.47)%,S期細(xì)胞比例分別為(25.40±0.42)%、(23.14±0.30)%、(20.90±0.25)%、(16.65±0.89)%、(9.41±0.73)%;統(tǒng)計(jì)結(jié)果顯示各組之間兩兩比較均有統(tǒng)計(jì)學(xué)差異(P0.05);說明TP可影響口腔鱗狀癌細(xì)胞的周期并有濃度依賴性。3.TRPCR結(jié)果顯示,24h后B、C、D、E組PTEN基因m RNA的表達(dá)率分別為對照組的1.67、2.22、2.86、3.5倍,72h后四組PTEN基因m RNA的表達(dá)率分別為對照組的3.95、5.27、7.18、9.16倍,各時(shí)間點(diǎn)的對照組與實(shí)驗(yàn)組以及各實(shí)驗(yàn)組組間兩兩比較均有統(tǒng)計(jì)學(xué)意義(P0.05);說明TP可上調(diào)PTEN中m RNA的表達(dá),并呈濃度依賴性。4.Western-blot結(jié)果顯示,TP作用口腔鱗狀癌細(xì)胞72h后,PTEN蛋白表達(dá)均上調(diào),分別為對照組的3.8倍、6.8倍、10.8倍、17.5倍,對照組和實(shí)驗(yàn)組以及各實(shí)驗(yàn)組間兩兩比較均有統(tǒng)計(jì)學(xué)差異(P0.05),說明了TP顯著上調(diào)了PTEN中蛋白的表達(dá),隨著TP濃度的增加,PTEN蛋白表達(dá)也呈現(xiàn)規(guī)律性增高。結(jié)論1在本實(shí)驗(yàn)條件下,TP可以有效抑制口腔鱗狀癌細(xì)胞的增殖;2 TP可將口腔鱗狀癌細(xì)胞抑制在G1期,減少了G1期向S期的轉(zhuǎn)變;3 TP可以提高口腔鱗狀癌細(xì)胞中PTEN的m RNA和蛋白的表達(dá)。
[Abstract]:Objective to investigate the effects of triptolide (TPTP) on the proliferation of human oral squamous cell carcinoma cell lines and the expression of homologous lost phosphatase (PTENs) on chromosome 10, and to explore the mechanism of TP in the treatment of oral squamous cell carcinoma (OSCC). To provide experimental evidence for the treatment of oral squamous cell carcinoma. Methods Oral squamous carcinoma cells were used as research objects. Oral squamous carcinoma cells were divided into two groups: control group (A) and four experimental groups. The cells were treated with TP of 0 1.25 渭 g / L of 2.55.0 and 10 渭 g / L respectively. 1. The increase of TP in oral squamous carcinoma cells was detected by methylthiazolyl tetrazolium methyl thiazolyl tetrazolium method MTTstaining after 24 h and 72 h. Effect of colonization changes on the effect of TP on the Cell cycle of Oral squamous carcinoma cells after 48 h Detection by flow Cytometry the effect of TP on the expression of Messenger RNA m RNA) in PTEN was detected by Real-Time Quantitative polymerase chain reactionation RT-PCR after 24 h and 72 h after Detection The effect of TP on the expression of PTEN protein was detected by Western-blot after 72 hours. Results (1) the cell inhibitory rates of MTT group were 12.48 鹵0.86 and 35.37 鹵0.105.37 鹵0.1010, respectively. The cell inhibition rates of the four experimental groups were 26.92 鹵0.14, 38.67 鹵0.112.62 鹵0.89, 90.42 鹵0.280.42 鹵0.280.42 鹵0.284h, respectively, compared with those of the control group, which were significantly higher than that of the control group (P 0.05H, P 0.05h), the cell inhibition rates of the four experimental groups were 26.92 鹵0.147.67 鹵0.112.62 鹵0.86.42 鹵0.281.Compared with the control group, the cell inhibition rates of the four experimental groups were significantly higher than those of the control group. The results showed that the cell inhibition rates of the four experimental groups were 26.92 鹵0.14, 38.67 鹵0.112.62 鹵0.89 and 90.42 鹵0.280.42 鹵0.28, respectively, compared with the control group. The cell inhibition rate increased with the increase of concentration, and the statistical analysis showed that there was a significant difference between the two groups, indicating that TP could inhibit the proliferation of oral squamous carcinoma cells. 2. The results of flow cytometry showed that the percentage of G 1 phase cells in the group treated with TP was 58.78 鹵0.98, 74.90 鹵0.281 鹵0.41 and 79.14 鹵0.455.The percentage of cells in S phase was 25.40 鹵0.42 and 20.90 鹵0.2565 鹵0.899.41 鹵0.899.41 鹵0.73.The statistical results showed that there was a statistical difference between the two groups, and the statistical results showed that there was a statistical difference between the two groups, indicating that there was a significant difference between the two groups in the percentage of cells in the G _ 1 phase, and the statistical results showed that there was a statistical difference between the two groups, indicating that there were significant differences between the two groups in the percentage of cells in the G _ 1 phase, and the statistical results showed that there was a significant difference between the two groups in the percentage of cells in the S phase, which was 25.40 鹵0.42 and 20.90 鹵0.259.65 鹵0.899.41 鹵0.73.The statistical results showed that there was a statistical difference between the two groups. TP could affect the cell cycle of oral squamous carcinoma cells in a concentration-dependent manner. The results of TRPCR showed that the expression rate of m RNA of PTEN gene in group E was 3.955.277.187.189.16 times higher than that in control group (1.67 2.222.863.5-fold) after 72 hours, and the expression rate of PTEN gene m RNA in the four groups was 3.955.277.181.16 times higher than that in the control group. The comparison between the control group and the experimental group at each time point had statistical significance (P 0.05), which indicated that TP could up-regulate the expression of m RNA in PTEN. In a concentration dependent manner, Western-blot showed that the expression of PTEN protein was up-regulated in oral squamous carcinoma cells 72 hours after treatment with TP, which was 6.8 times, 10.8 times and 17.5 times as much as that of the control group, respectively. There was a significant difference between the control group and the experimental group (P 0.05), which indicated that TP upregulated the expression of protein in PTEN, and the expression of PTEN increased regularly with the increase of TP concentration. Conclusion (1) under this condition, TP can effectively inhibit the proliferation of oral squamous carcinoma cells and can inhibit the proliferation of oral squamous carcinoma cells in G1 phase. The expression of m RNA and protein of PTEN in oral squamous carcinoma cells was increased by decreasing the transformation from G1 phase to S phase.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R739.8

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 王宏偉;程小麗;許雅清;邱家權(quán);李海龍;吳玉泓;徐燕;;苦馬豆素對胃癌細(xì)胞中pten和survivin mRNA表達(dá)的影響[J];西部中醫(yī)藥;2015年12期

2 卜強(qiáng);曾明輝;蔣華;蔣東方;秦鎖炳;陳巧云;李皓;;吡柔比星聯(lián)合雷公藤甲素對人膀胱癌T24細(xì)胞生長的影響[J];腫瘤;2014年06期

3 趙紅霞;;PTEN與腫瘤的相關(guān)性研究進(jìn)展[J];武漢大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2014年01期

4 孟令飛;李蔚;王翠;;抑癌基因PTEN在人類惡性腫瘤中的表達(dá)及其研究進(jìn)展[J];中國冶金工業(yè)醫(yī)學(xué)雜志;2013年04期

5 孫曉菊;左海燕;謝洪;鄧巍;;口腔鱗狀細(xì)胞癌中PTEN、MMP-7和VEGF的表達(dá)及相關(guān)性研究[J];中國實(shí)用口腔科雜志;2013年03期

6 郭偉健;何敏毅;孫學(xué)剛;劉亞偉;彭康;;雷公藤甲素對大腸癌細(xì)胞細(xì)胞周期的調(diào)控作用[J];貴陽醫(yī)學(xué)院學(xué)報(bào);2012年02期

7 夏旭芬;王偉;鮑亞萍;張偉敏;姚航平;童向民;;雷公藤內(nèi)酯醇對結(jié)腸癌細(xì)胞COX-2和iNOS表達(dá)的抑制作用[J];中國藥學(xué)雜志;2008年10期

8 王國平;尹成進(jìn);歐陽曙明;鄭乃青;楊業(yè)金;李文庭;;雷公藤內(nèi)酯醇對人胃癌細(xì)胞SGC7901增殖及表達(dá)血管內(nèi)皮生長因子的影響[J];實(shí)用醫(yī)學(xué)雜志;2008年01期

9 鄭家偉;李金忠;鐘來平;張志愿;;口腔鱗狀細(xì)胞癌臨床流行病學(xué)研究現(xiàn)狀[J];中國口腔頜面外科雜志;2007年02期

10 呂秀紅;郭瑞珍;刁路明;劉銘球;;雷公藤內(nèi)酯醇對肺腺癌細(xì)胞系A(chǔ)549的體外抑制作用的研究[J];中國藥理學(xué)通報(bào);2007年02期

相關(guān)博士學(xué)位論文 前3條

1 姜寧;雷公藤甲素對人肺腺癌紫杉醇耐藥細(xì)胞A549/Taxol的耐藥逆轉(zhuǎn)作用及機(jī)制[D];山東大學(xué);2016年

2 楊思俊;癌基因RMP與抑癌基因PTEN的相互拮抗作用及其對肝癌細(xì)胞增殖的影響[D];蘇州大學(xué);2015年

3 齊世美;磷酸化熱休克蛋白27與腫瘤細(xì)胞TRAIL耐受的分子機(jī)制的研究[D];南京師范大學(xué);2012年

相關(guān)碩士學(xué)位論文 前1條

1 曾夏梅;口腔白斑及鱗癌中PCNA、p27、PTEN基因的表達(dá)[D];福建醫(yī)科大學(xué);2012年

，

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