本文選題：生物鐘 + 癌　； 參考：《重慶醫(yī)科大學》2017年碩士論文

【摘要】：目的:探討口腔鱗癌細胞內(nèi)生物鐘基因PER1對生物鐘基因網(wǎng)絡中其它生物鐘基因表達的影響。方法:通過RNA干擾技術沉默PER1并構(gòu)建3組PER1短發(fā)夾(short hairpin)RNA慢病毒載體質(zhì)粒,然后分別感染SCC15口腔鱗癌細胞株,經(jīng)Western blot和實時熒光定量PCR(Quantitative real-time PCR,qRT-PCR)篩選并確定PER1沉默效果最佳組定為實驗組,以轉(zhuǎn)染scramble質(zhì)粒(不含任何干擾基因片段)的SCC15細胞為陰性對照組,以相同培養(yǎng)條件下不曾做處理的SCC15細胞為空白對照組。分別在體外和體內(nèi)應用qRT-PCR檢測PER1沉默前后生物鐘基因CLOCK、BMAL1、PER2、PER3、DEC1、DEC2、CRY1、CRY2、TIM、CKIε、RORα、NPAS2和REV-ERBαmRNA的表達情況,應用流式細胞儀檢測口腔鱗癌細胞的增殖和凋亡水平的改變情況,并應用裸鼠體內(nèi)成瘤實驗觀察SCC15癌細胞體內(nèi)成瘤能力改變情況。結(jié)果:PER1基因沉默后,體內(nèi)和體外癌細胞中生物鐘基因PER2、DEC1、DEC2、CRY1、CRY2和NPAS2 mRNA的表達顯著降低(P0.05),PER3、TIM、RORɑ和REV-ERBɑmRNA的表達顯著升高(P0.05),而CLOCK、BMAL1和CKIεmRNA的表達無顯著改變(P0.05)。PER1沉默之后,SCC15癌細胞的凋亡降低,增殖和體內(nèi)成瘤加強。結(jié)論:生物鐘基因PER1可以調(diào)控鐘基因網(wǎng)絡中眾多其他鐘基因如PER2、DEC1、DEC2、CRY1、CRY2、NPAS2、PER3、TIM、RORɑ和REV-ERBɑ的表達,PER1在機體的鐘基因網(wǎng)絡中具有十分重要的調(diào)控功能。PER1對癌癥形成的作用并非單一通過對其下游基因的調(diào)控,而是同時通過對生物鐘基因網(wǎng)絡中諸多生物鐘基因的異常調(diào)控所共同導致。
[Abstract]:Aim: to investigate the effect of clock gene PER1 on the expression of other clock genes in oral squamous cell carcinoma (OSCC) cells. Methods: PER1 was silenced by RNA interference technique and three groups of PER1 short hairpin)RNA vector plasmids were constructed and then infected with SCC15 oral squamous cell carcinoma cell lines. Western blot and real-time quantitative PCR(Quantitative real-time PCR qRT-PCR were used to screen and determine the best effect of PER1 silencing. SCC15 cells transfected with scramble plasmid (without any interfering gene fragments) were used as negative control group. The SCC15 cells which were not treated under the same culture conditions were used as the blank control group. In vitro and in vivo, qRT-PCR was used to detect the expression of circadian clock gene CLOCKC BMAL1, PER2PER3, DEC1, CRY2, CRY2, CRY2, CRY2, and REV-ERB 偽 mRNA, and flow cytometry was used to detect the changes of proliferation and apoptosis in oral squamous carcinoma cells, respectively, before and after PER1 silencing, and in vitro and in vivo, respectively, were used to detect the expression of REV-ERB 偽 -NPAS2 and REV-ERB 偽 mRNA in oral squamous carcinoma cells, respectively, before and after PER1 silencing. The tumorigenic ability of SCC15 cancer cells in vivo was observed by tumorigenesis test in nude mice. Results the expression of clock gene PER2 / DEC1 / CRY1CRY2 and NPAS2 mRNA in cancer cells decreased significantly after the gene was silenced. The expression of P0.05, PER3TIMROR and REV-ERB mRNA increased significantly, while the expression of CLO CKBMAL1 and CKI 蔚 mRNA did not change the apoptosis of SCC15 cells. Proliferation and tumorigenesis are enhanced. Conclusion: biological clock gene PER1 can regulate the expression of many other clock genes in the clock gene network, such as PER2DEC1, DEC1, CRY1, CRY2, NPAS2, PER3, TIMROR and REV-ERB. PER1 plays a very important regulatory role in the clock gene network. PER1 plays an important role in cancer formation. The regulation of its downstream genes, At the same time, it is caused by the abnormal regulation of many biological clock genes in the biological clock gene network.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R739.8

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相關期刊論文 前2條

1 李晗雪;楊凱;付小娟;趙欽;;生物鐘基因Per1對人口腔鱗癌細胞生物學行為的影響和調(diào)控機制[J];中國醫(yī)學科學院學報;2016年02期

2 葉華;楊凱;譚雪梅;趙丹;呂曉強;王青青;;鐘基因Per2和腫瘤相關鐘控基因在金黃地鼠口腔頰黏膜癌變不同階段的晝夜節(jié)律改變[J];華西口腔醫(yī)學雜志;2015年05期

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本文編號：1846186