本文選題：口腔鱗狀細胞癌 + 雷公藤屬��； 參考：《華北理工大學》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)基因表達的影響,探討TP抗口腔鱗狀癌的作用機制,以期為口腔鱗狀細胞癌的治療提供實驗依據(jù)。方法以口腔鱗狀癌細胞作為研究對象,將口腔鱗狀癌細胞分為對照組(A)和四個實驗組(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對口腔鱗狀癌細胞增殖變化的影響;2流式細胞檢測技術檢測48h后TP對口腔鱗狀癌細胞周期的影響;3實時熒光定量(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)表達的影響;4蛋白印跡法(Western-blot)檢測72h后TP對PTEN蛋白表達的影響。結果1 MTT結果:24h后B、C、D、E組的細胞抑制率分別為(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)計學意義(P0.05);72h后四個實驗組的細胞抑制率分別為(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)計學意義(P0.05);48h后,細胞抑制率隨濃度增加而增加,統(tǒng)計分析結果顯示各組間兩兩比較均有統(tǒng)計學意義(P0.05);說明TP可抑制口腔鱗狀癌細胞增殖。2.流式細胞檢測技術結果顯示,TP作用口腔鱗狀癌細胞48后,A、B、C、D、E各組G1期細胞比例分別為(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)計結果顯示各組之間兩兩比較均有統(tǒng)計學差異(P0.05);說明TP可影響口腔鱗狀癌細胞的周期并有濃度依賴性。3.TRPCR結果顯示,24h后B、C、D、E組PTEN基因m RNA的表達率分別為對照組的1.67、2.22、2.86、3.5倍,72h后四組PTEN基因m RNA的表達率分別為對照組的3.95、5.27、7.18、9.16倍,各時間點的對照組與實驗組以及各實驗組組間兩兩比較均有統(tǒng)計學意義(P0.05);說明TP可上調PTEN中m RNA的表達,并呈濃度依賴性。4.Western-blot結果顯示,TP作用口腔鱗狀癌細胞72h后,PTEN蛋白表達均上調,分別為對照組的3.8倍、6.8倍、10.8倍、17.5倍,對照組和實驗組以及各實驗組間兩兩比較均有統(tǒng)計學差異(P0.05),說明了TP顯著上調了PTEN中蛋白的表達,隨著TP濃度的增加,PTEN蛋白表達也呈現(xiàn)規(guī)律性增高。結論1在本實驗條件下,TP可以有效抑制口腔鱗狀癌細胞的增殖;2 TP可將口腔鱗狀癌細胞抑制在G1期,減少了G1期向S期的轉變;3 TP可以提高口腔鱗狀癌細胞中PTEN的m RNA和蛋白的表達。
[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.
【學位授予單位】：華北理工大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R739.8

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