多氯聯(lián)苯醌誘導(dǎo)細(xì)胞發(fā)生自噬以及自噬—凋亡轉(zhuǎn)換的機(jī)制分析
發(fā)布時(shí)間：2018-07-25 19:32
【摘要】：多氯聯(lián)苯(PCBs)是一類無(wú)所不在的持久性有機(jī)污染物,由于其多種毒性效應(yīng)如內(nèi)分泌毒性、免疫毒性、神經(jīng)毒性等,在二十世紀(jì)七十年代已被禁止使用,但是由于其特殊的理化及生物學(xué)特性,PCBs仍然廣泛存在于我們的環(huán)境包括陸地和水生系統(tǒng)。細(xì)胞自噬是細(xì)胞內(nèi)受損衰老的蛋白質(zhì)或者細(xì)胞器包裹起來(lái)形成自噬體,自噬體再與溶酶體融合進(jìn)而進(jìn)行消化降解的過(guò)程,也常被作為細(xì)胞的自我保護(hù)機(jī)制。該研究的目的是為了分析PCB29-pQ激活自噬的具體機(jī)制,以及探討細(xì)胞自噬與細(xì)胞凋亡間的轉(zhuǎn)換機(jī)制。第一部分:該部分研究考察了多氯聯(lián)苯醌通過(guò)mTOR/p70S6k誘導(dǎo)細(xì)胞自噬發(fā)生的分子機(jī)制。選用HepG2和MDA-MB-231細(xì)胞作為研究對(duì)象,并通過(guò)實(shí)驗(yàn)發(fā)現(xiàn)PCB29-pQ誘導(dǎo)的自噬沒(méi)有細(xì)胞特異性。首先,兩種細(xì)胞在5μM PCB29-pQ處理24 h后透射電鏡檢測(cè)細(xì)胞超微結(jié)構(gòu)顯示出自噬泡顯著特征,并通過(guò)AO染色、MDC染色觀察到明顯的自噬泡形成,由此先從形態(tài)學(xué)和生化特征表明PCB29-pQ作用后可以引起細(xì)胞發(fā)生自噬。然后從自噬體形成的3個(gè)階段證明誘導(dǎo)自噬發(fā)生的分子機(jī)制。第一階段從濃度和時(shí)間梯度上分別檢測(cè)了對(duì)自噬開(kāi)關(guān)負(fù)調(diào)控的蛋白mTOR和p70S6k的表達(dá),在PCB29-pQ作用后其蛋白表達(dá)水平降低,表明激活了自噬的誘導(dǎo)階段。第二階段檢測(cè)了自噬體形成過(guò)程中自噬相關(guān)蛋白ATG5、ATG12、LC3的表達(dá),并用RT-PCR分析LC3 mRNA水平。與對(duì)照組相比,PCB29-pQ處理的細(xì)胞蛋白和基因的表達(dá)水平都顯著增加,且在5μM PCB29-pQ處理24 h有最大值。而自噬降解底物p62蛋白水平隨時(shí)間梯度降低,在5μM PCB29-pQ處理24 h表達(dá)水平最低。這些結(jié)果說(shuō)明PCB29-pQ能夠激活自噬體的形成階段,促進(jìn)自噬體降解底物的能力。第三階段通過(guò)加入CQ抑制LC3的降解后檢測(cè)LC3表達(dá)的凈通量,結(jié)果表明PCB29-pQ作用后提高了LC3凈通量并在5μM PCB29-pQ作用24 h時(shí)最明顯。并且通過(guò)轉(zhuǎn)染GFP-LC3質(zhì)粒,免疫熒光觀察了自噬體標(biāo)記LC3與溶酶體標(biāo)記LAMP2的共定位,結(jié)果表明促進(jìn)了自噬體與溶酶體融合的階段。接著闡明了自噬的發(fā)生在PCB29-pQ誘導(dǎo)細(xì)胞毒性中的作用,在加入不同階段自噬抑制劑3-MA和CQ后,通過(guò)CCK8、流式細(xì)胞儀、細(xì)胞凋亡標(biāo)志蛋白caspase3的檢測(cè),表明抑制自噬促進(jìn)了PCB29-pQ誘導(dǎo)的細(xì)胞毒性的增加。最后用加入ROS清除劑NAC的方法考察PCB29-pQ引起自噬水平升高的原因,結(jié)果表明,PCB29-pQ誘導(dǎo)的細(xì)胞自噬的激活受ROS水平的調(diào)節(jié)。根據(jù)以上研究可知,PCB29-pQ可以誘導(dǎo)HepG2和MDA-MB-231細(xì)胞發(fā)生自噬,并受細(xì)胞內(nèi)ROS水平的調(diào)節(jié)。PCB29-pQ引起的自噬受mTOR/p70S6k和ATG5/ATG12/LC3信號(hào)通路的調(diào)控,且作為一種存活機(jī)制保護(hù)細(xì)胞。第二部分:該部分初步探討了多氯聯(lián)苯醌在低濃度誘導(dǎo)自噬,高濃度誘導(dǎo)凋亡時(shí),鈣蛋白酶活性的作用對(duì)自噬向凋亡信號(hào)傳遞的影響。首先通過(guò)JC-1熒光探針檢測(cè)HepG2細(xì)胞線粒體膜電位,免疫印跡法檢測(cè)凋亡相關(guān)蛋白caspase9/caspase3等表達(dá),TUNEL法檢測(cè)細(xì)胞凋亡,免疫熒光檢測(cè)自噬標(biāo)志蛋白LC3焦點(diǎn),結(jié)果發(fā)現(xiàn),與對(duì)照組相比,隨著PCB29-pQ處理濃度的升高細(xì)胞凋亡水平逐漸增加,而在低濃度5μM PCB29-pQ處理后自噬LC3焦點(diǎn)最多,隨著濃度升高,自噬LC3焦點(diǎn)數(shù)目降低。隨后用流式細(xì)胞儀檢測(cè)PCB29-pQ誘導(dǎo)的細(xì)胞鈣離子水平的變化,熒光分光光度計(jì)檢測(cè)鈣蛋白酶活性,結(jié)果表明,PCB29-pQ可以誘導(dǎo)HepG2細(xì)胞鈣離子水平以及鈣蛋白酶活性的升高,以及在加入鈣離子螯合劑BAPTA-AM后抑制了calpain蛋白表達(dá),表明,calpain的表達(dá)依賴于Ca2+水平的調(diào)節(jié)。文獻(xiàn)報(bào)道Beclin1、ATG5可經(jīng)鈣蛋白酶切割形成Beclin1-c、tATG5易位至線粒體將自噬向凋亡信號(hào)傳遞,而在本研究中PCB29-pQ沒(méi)有引起B(yǎng)eclin1、ATG5切割易位至線粒體。以上研究可知,PCB29-pQ可以在低濃度時(shí)誘導(dǎo)HepG2細(xì)胞自噬,高濃度時(shí)誘導(dǎo)細(xì)胞凋亡,并且伴隨著細(xì)胞內(nèi)鈣離子水平和鈣蛋白酶活性的升高,但鈣蛋白酶活性并不引起B(yǎng)eclin1、ATG5產(chǎn)生切割將信號(hào)從自噬傳遞至凋亡。第三部分:該部分進(jìn)一步探討了多氯聯(lián)苯醌誘導(dǎo)細(xì)胞自噬與細(xì)胞凋亡轉(zhuǎn)換的機(jī)制,分析選取了對(duì)自噬和凋亡有雙重調(diào)節(jié)作用的p53/HMGB1蛋白。Western Blot和免疫熒光實(shí)驗(yàn)表明了PCB29-pQ作用后誘導(dǎo)HepG2細(xì)胞p53/HMGB1易位到細(xì)胞質(zhì)。免疫共沉淀實(shí)驗(yàn)表明了5μM PCB29-pQ作用后促進(jìn)了HepG2細(xì)胞p53/HMGB1在細(xì)胞核中的結(jié)合,15μM PCB29-pQ作用后促進(jìn)了HepG2細(xì)胞p53/HMGB1在細(xì)胞質(zhì)中的結(jié)合。由于HMGB1/p53可以作為核轉(zhuǎn)錄因子調(diào)控下游靶基因的表達(dá),影響細(xì)胞自噬與凋亡。我們干擾了p53蛋白表達(dá)后檢測(cè)了p53下游靶基因DRAM、ULK1、Bax表達(dá),同時(shí)干擾了HMGB1蛋白表達(dá)后Western Blot檢測(cè)了HMGB1的下游蛋白HSPB1表達(dá),結(jié)果表明,在抑制了p53/HMGB1的作用后,抑制了該蛋白調(diào)控的靶基因的表達(dá)。針對(duì)上述檢測(cè)結(jié)果,我們進(jìn)一步研究p53/HMGB1誘導(dǎo)細(xì)胞自噬和細(xì)胞凋亡的作用機(jī)制。干擾p53基因后提取了核質(zhì)蛋白,Western Blot表明p53 siRNA后促進(jìn)了PCB29-pQ誘導(dǎo)的HMGB1進(jìn)一步易位至細(xì)胞質(zhì),細(xì)胞存活率實(shí)驗(yàn)表明了p53 siRNA抑制了PCB29-pQ誘導(dǎo)的細(xì)胞凋亡,免疫熒光LC3焦點(diǎn)實(shí)驗(yàn)表明p53 siRNA促進(jìn)了PCB29-pQ誘導(dǎo)的細(xì)胞自噬,而在加入HMGB1抑制劑EP后,細(xì)胞凋亡增加,細(xì)胞自噬水平降低。這些結(jié)論表明,易位到細(xì)胞質(zhì)中的HMGB1發(fā)揮著抑制凋亡、促進(jìn)自噬的作用。在干擾了HMGB1蛋白后相同的實(shí)驗(yàn)方法證明HMGB1 siRNA后促進(jìn)了PCB29-pQ誘導(dǎo)的p53進(jìn)一步易位至細(xì)胞質(zhì),促進(jìn)了PCB29-pQ誘導(dǎo)的細(xì)胞凋亡,以及抑制了PCB29-pQ誘導(dǎo)的細(xì)胞自噬,而在加入p53抑制劑PFT-α后,細(xì)胞凋亡減少,細(xì)胞自噬水平升高。這些結(jié)論表明,易位到細(xì)胞質(zhì)中的p53發(fā)揮著促進(jìn)凋亡、抑制自噬的作用。以上研究表明,PCB29-pQ誘導(dǎo)HepG2細(xì)胞自噬與細(xì)胞凋亡的過(guò)程中,是通過(guò)p53/HMGB1在細(xì)胞核與細(xì)胞質(zhì)定位以及對(duì)靶基因表達(dá)的不同發(fā)揮其在PCB29-pQ引起細(xì)胞自噬與細(xì)胞凋亡間的調(diào)控作用。
[Abstract]:Polychlorinated biphenyl (PCBs) is a ubiquitous persistent organic pollutant. Due to its many toxic effects, such as endocrine toxicity, immunotoxicity, neurotoxicity, and so on, it has been banned in 1970s, but because of its special physicochemical and biological characteristics, PCBs still exists in our environment including land and aquatic products. Autophagy is the process of cell autophagy or cell organelle wrapped up to form autophagosome, and the autophagosome is fused with lysosomes for digestion and degradation, and is often used as a self-protection mechanism for cells. The purpose of this study is to analyze the specific mechanism of PCB29-pQ activation and to explore the autophagy of cells. The transformation mechanism with cell apoptosis. Part 1: this part studied the molecular mechanism of polychlorinated biphenyquinone induced autophagy through mTOR/p70S6k. HepG2 and MDA-MB-231 cells were selected as the research object, and the experiment found that the autophagy induced by PCB29-pQ had no cell specificity. First, two cells were treated in 5 mu M PCB29-pQ. After 24 h, the ultrastructure of the cell was detected by the transmission electron microscope. The obvious autophagy was observed by AO staining and MDC staining was used to observe the formation of autophagic vesicles. From the morphological and biochemical characteristics, the cell autophagy could be caused by the action of PCB29-pQ. Then, the molecular mechanism of inducing autophagy was proved from the 3 stages of autophagic formation. In the first stage, the expression of protein mTOR and p70S6k negatively regulated by autophagic switch was detected from the concentration and time gradient, and the expression level of the protein decreased after the action of PCB29-pQ, indicating the activation of the induction stage of autophagy. The second stage detected the expression of autophagic related protein ATG5, ATG12, LC3 and RT-PCR in the process of autophagic formation. Analysis of LC3 mRNA level. Compared with the control group, the expression level of cell protein and gene in PCB29-pQ treatment increased significantly, and the maximum value was 24 h at 5 M PCB29-pQ treatment. The level of the autophagy degradation substrate p62 protein decreased with the time gradient, and the lowest expression level in the 5 u M PCB29-pQ processing 24 h. These results indicate that PCB29-pQ can activate autophagy. The formation stage of the body promotes the ability of the autophagic to degrade the substrate. The third phase detects the net flux of LC3 expression by adding CQ to inhibit the degradation of LC3. The results show that the net flux of LC3 is enhanced after the action of PCB29-pQ and is most obvious at the action of 24 h by 5 M PCB29-pQ. And the expression of the autophagic marker LC3 is observed by the transfection of the GFP-LC3 particles. The co localization of lysosome labeled LAMP2 showed that the fusion of autophagosomes and lysosomes was promoted. Then the role of autophagy in PCB29-pQ induced cytotoxicity was clarified. After adding different stages of autophagy inhibitor 3-MA and CQ, the detection of the inhibition of autophagy through the detection of CCK8, flow cytometry, and apoptosis marker protein Caspase3. The increase of cytotoxicity induced by PCB29-pQ was promoted. Finally, the reasons for the increase of autophagy induced by PCB29-pQ were investigated by the addition of ROS scavenger NAC. The results showed that the activation of autophagy induced by PCB29-pQ was regulated by the ROS level. According to the above study, PCB29-pQ could induce autophagy in HepG2 and MDA-MB-231 cells and was finer. The autophagy induced by the intracellular ROS level of.PCB29-pQ is regulated by the mTOR/p70S6k and ATG5/ATG12/LC3 signaling pathways and protects the cells as a survival mechanism. The second part: this part preliminarily discussed the effect of the activity of calcium protease on the autophagy to the apoptosis signal when the autophagy induced by the low concentration of polychlorinated biphenyquinone. The JC-1 fluorescence probe was used to detect the mitochondrial membrane potential of HepG2 cells, and the expression of apoptosis related protein caspase9/caspase3 was detected by immunoblotting. Apoptosis was detected by TUNEL method and the focal point of autophagic marker protein LC3 was detected by immunofluorescence. The results were found to be compared with the control group, and the apoptotic water was increased with the concentration of PCB29-pQ. The level of autophagy was increased gradually, and the focus of autophagy LC3 was most in the low concentration of 5 M PCB29-pQ. With the increase of concentration, the number of autophagic LC3 focal points decreased. Then the level of calcium ion induced by PCB29-pQ was detected by flow cytometry, and the activity of calsin was detected by the fluorescence spectrophotometer. The results showed that PCB29-pQ could induce the calcium ion of HepG2 cells. Levels and the increase of calsin activity and the inhibition of calpain protein expression after adding calcium ion chelating agent BAPTA-AM showed that the expression of calpain depended on the regulation of Ca2+ level. The literature reported that Beclin1, ATG5 can be cut through calsin to form Beclin1-c, tATG5 translocation to mitochondria to transmit autophagy to apoptosis signal, and in this study PCB29-pQ did not cause Beclin1, ATG5 cut translocation to mitochondria. Above study, PCB29-pQ can induce autophagy at low concentration of HepG2 cells, induce apoptosis at high concentration, and increase the intracellular calcium level and calsin activity, but the activity of calcinease does not cause Beclin1, ATG5 produces cutting signal. From autophagy to apoptosis. The third part: this part further explored the mechanism of polychlorinated biphenyquinone induced autophagy and cell apoptosis, and selected the p53/HMGB1 protein.Western Blot and immunofluorescence test, which had dual regulation on autophagy and apoptosis, and showed that PCB29-pQ could induce p53/HMGB1 translocation to HepG2 cells after PCB29-pQ action. Cytoplasm. The immunoprecipitation experiment showed that the interaction of 5 M PCB29-pQ promoted the binding of p53/HMGB1 in the nucleus and promoted the binding of p53/HMGB1 in the cytoplasm after the action of 15 mu M PCB29-pQ. Because HMGB1/p53 could be used as a nuclear transcription factor to regulate the expression of the target gene of the downstream target and affect the autophagy and apoptosis of the cell. After interfering with the expression of p53 protein, the downstream target gene of p53 was detected, DRAM, ULK1, Bax expression, and Western Blot detected the downstream protein HSPB1 expression of HMGB1 after the expression of HMGB1 protein. The results showed that the expression of the target gene was suppressed after the inhibition of p53/HMGB1 action. We further studied the results of the above detection. The mechanism of p53/HMGB1 induced autophagy and apoptosis of cells. After interfering with the p53 gene, the nuclear protein was extracted, and Western Blot showed that p53 siRNA promoted the PCB29-pQ induced HMGB1 to be further translocated to the cytoplasm. The cell survival rate experiment showed that p53 siRNA inhibited the apoptosis of PCB29-pQ induced cells, and the immunofluorescent LC3 focus experiment table P53 siRNA promoted the autophagy induced by PCB29-pQ, and the apoptosis increased and the level of autophagy decreased after the addition of HMGB1 inhibitor EP. These conclusions suggest that the HMGB1 in the cytoplasm may play a role in inhibiting apoptosis and promoting autophagy. After the interference of HMGB1 protein, the same experimental method proved that HMGB1 siRNA promoted PCB2. 9-pQ induced p53 further translocation to cytoplasm, promoting apoptosis induced by PCB29-pQ and inhibiting autophagy induced by PCB29-pQ, and the decrease of cell apoptosis and the increase of autophagy after adding the p53 inhibitor PFT- alpha. These conclusions suggest that p53 in the translocation of the cytoplasm plays a role in promoting apoptosis and inhibiting autophagy. The study shows that PCB29-pQ induces autophagy and apoptosis in HepG2 cells, which regulates the role of p53/HMGB1 in cell autophagy and cell apoptosis through the localization of the nucleus and cytoplasm and the difference in the expression of the target gene.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R114
本文編號(hào)：2144848
[Abstract]:Polychlorinated biphenyl (PCBs) is a ubiquitous persistent organic pollutant. Due to its many toxic effects, such as endocrine toxicity, immunotoxicity, neurotoxicity, and so on, it has been banned in 1970s, but because of its special physicochemical and biological characteristics, PCBs still exists in our environment including land and aquatic products. Autophagy is the process of cell autophagy or cell organelle wrapped up to form autophagosome, and the autophagosome is fused with lysosomes for digestion and degradation, and is often used as a self-protection mechanism for cells. The purpose of this study is to analyze the specific mechanism of PCB29-pQ activation and to explore the autophagy of cells. The transformation mechanism with cell apoptosis. Part 1: this part studied the molecular mechanism of polychlorinated biphenyquinone induced autophagy through mTOR/p70S6k. HepG2 and MDA-MB-231 cells were selected as the research object, and the experiment found that the autophagy induced by PCB29-pQ had no cell specificity. First, two cells were treated in 5 mu M PCB29-pQ. After 24 h, the ultrastructure of the cell was detected by the transmission electron microscope. The obvious autophagy was observed by AO staining and MDC staining was used to observe the formation of autophagic vesicles. From the morphological and biochemical characteristics, the cell autophagy could be caused by the action of PCB29-pQ. Then, the molecular mechanism of inducing autophagy was proved from the 3 stages of autophagic formation. In the first stage, the expression of protein mTOR and p70S6k negatively regulated by autophagic switch was detected from the concentration and time gradient, and the expression level of the protein decreased after the action of PCB29-pQ, indicating the activation of the induction stage of autophagy. The second stage detected the expression of autophagic related protein ATG5, ATG12, LC3 and RT-PCR in the process of autophagic formation. Analysis of LC3 mRNA level. Compared with the control group, the expression level of cell protein and gene in PCB29-pQ treatment increased significantly, and the maximum value was 24 h at 5 M PCB29-pQ treatment. The level of the autophagy degradation substrate p62 protein decreased with the time gradient, and the lowest expression level in the 5 u M PCB29-pQ processing 24 h. These results indicate that PCB29-pQ can activate autophagy. The formation stage of the body promotes the ability of the autophagic to degrade the substrate. The third phase detects the net flux of LC3 expression by adding CQ to inhibit the degradation of LC3. The results show that the net flux of LC3 is enhanced after the action of PCB29-pQ and is most obvious at the action of 24 h by 5 M PCB29-pQ. And the expression of the autophagic marker LC3 is observed by the transfection of the GFP-LC3 particles. The co localization of lysosome labeled LAMP2 showed that the fusion of autophagosomes and lysosomes was promoted. Then the role of autophagy in PCB29-pQ induced cytotoxicity was clarified. After adding different stages of autophagy inhibitor 3-MA and CQ, the detection of the inhibition of autophagy through the detection of CCK8, flow cytometry, and apoptosis marker protein Caspase3. The increase of cytotoxicity induced by PCB29-pQ was promoted. Finally, the reasons for the increase of autophagy induced by PCB29-pQ were investigated by the addition of ROS scavenger NAC. The results showed that the activation of autophagy induced by PCB29-pQ was regulated by the ROS level. According to the above study, PCB29-pQ could induce autophagy in HepG2 and MDA-MB-231 cells and was finer. The autophagy induced by the intracellular ROS level of.PCB29-pQ is regulated by the mTOR/p70S6k and ATG5/ATG12/LC3 signaling pathways and protects the cells as a survival mechanism. The second part: this part preliminarily discussed the effect of the activity of calcium protease on the autophagy to the apoptosis signal when the autophagy induced by the low concentration of polychlorinated biphenyquinone. The JC-1 fluorescence probe was used to detect the mitochondrial membrane potential of HepG2 cells, and the expression of apoptosis related protein caspase9/caspase3 was detected by immunoblotting. Apoptosis was detected by TUNEL method and the focal point of autophagic marker protein LC3 was detected by immunofluorescence. The results were found to be compared with the control group, and the apoptotic water was increased with the concentration of PCB29-pQ. The level of autophagy was increased gradually, and the focus of autophagy LC3 was most in the low concentration of 5 M PCB29-pQ. With the increase of concentration, the number of autophagic LC3 focal points decreased. Then the level of calcium ion induced by PCB29-pQ was detected by flow cytometry, and the activity of calsin was detected by the fluorescence spectrophotometer. The results showed that PCB29-pQ could induce the calcium ion of HepG2 cells. Levels and the increase of calsin activity and the inhibition of calpain protein expression after adding calcium ion chelating agent BAPTA-AM showed that the expression of calpain depended on the regulation of Ca2+ level. The literature reported that Beclin1, ATG5 can be cut through calsin to form Beclin1-c, tATG5 translocation to mitochondria to transmit autophagy to apoptosis signal, and in this study PCB29-pQ did not cause Beclin1, ATG5 cut translocation to mitochondria. Above study, PCB29-pQ can induce autophagy at low concentration of HepG2 cells, induce apoptosis at high concentration, and increase the intracellular calcium level and calsin activity, but the activity of calcinease does not cause Beclin1, ATG5 produces cutting signal. From autophagy to apoptosis. The third part: this part further explored the mechanism of polychlorinated biphenyquinone induced autophagy and cell apoptosis, and selected the p53/HMGB1 protein.Western Blot and immunofluorescence test, which had dual regulation on autophagy and apoptosis, and showed that PCB29-pQ could induce p53/HMGB1 translocation to HepG2 cells after PCB29-pQ action. Cytoplasm. The immunoprecipitation experiment showed that the interaction of 5 M PCB29-pQ promoted the binding of p53/HMGB1 in the nucleus and promoted the binding of p53/HMGB1 in the cytoplasm after the action of 15 mu M PCB29-pQ. Because HMGB1/p53 could be used as a nuclear transcription factor to regulate the expression of the target gene of the downstream target and affect the autophagy and apoptosis of the cell. After interfering with the expression of p53 protein, the downstream target gene of p53 was detected, DRAM, ULK1, Bax expression, and Western Blot detected the downstream protein HSPB1 expression of HMGB1 after the expression of HMGB1 protein. The results showed that the expression of the target gene was suppressed after the inhibition of p53/HMGB1 action. We further studied the results of the above detection. The mechanism of p53/HMGB1 induced autophagy and apoptosis of cells. After interfering with the p53 gene, the nuclear protein was extracted, and Western Blot showed that p53 siRNA promoted the PCB29-pQ induced HMGB1 to be further translocated to the cytoplasm. The cell survival rate experiment showed that p53 siRNA inhibited the apoptosis of PCB29-pQ induced cells, and the immunofluorescent LC3 focus experiment table P53 siRNA promoted the autophagy induced by PCB29-pQ, and the apoptosis increased and the level of autophagy decreased after the addition of HMGB1 inhibitor EP. These conclusions suggest that the HMGB1 in the cytoplasm may play a role in inhibiting apoptosis and promoting autophagy. After the interference of HMGB1 protein, the same experimental method proved that HMGB1 siRNA promoted PCB2. 9-pQ induced p53 further translocation to cytoplasm, promoting apoptosis induced by PCB29-pQ and inhibiting autophagy induced by PCB29-pQ, and the decrease of cell apoptosis and the increase of autophagy after adding the p53 inhibitor PFT- alpha. These conclusions suggest that p53 in the translocation of the cytoplasm plays a role in promoting apoptosis and inhibiting autophagy. The study shows that PCB29-pQ induces autophagy and apoptosis in HepG2 cells, which regulates the role of p53/HMGB1 in cell autophagy and cell apoptosis through the localization of the nucleus and cytoplasm and the difference in the expression of the target gene.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R114
【參考文獻(xiàn)】
相關(guān)期刊論文 前5條
1 王寵;張萍;朱衛(wèi)國(guó);;細(xì)胞自噬與腫瘤發(fā)生的關(guān)系[J];中國(guó)生物化學(xué)與分子生物學(xué)報(bào);2010年11期
2 曹先仲;陳花果;申松梅;宋艷輝;連小英;;多氯聯(lián)苯的性質(zhì)及其對(duì)環(huán)境的危害[J];中國(guó)科技論文在線;2008年05期
3 楊紹杰;孟金萍;屈yN;劉云波;;細(xì)胞凋亡信號(hào)傳導(dǎo)通路的研究進(jìn)展[J];中國(guó)比較醫(yī)學(xué)雜志;2007年05期
4 龔鐘明,曹軍,李本綱,徐福留,陶澍,沈偉然,張文具,韓寶平,孫韌;天津地區(qū)土壤中六六六(HCH)的殘留及分布特征[J];中國(guó)環(huán)境科學(xué);2003年03期
5 孫士勇,韓銳;活性氧與腫瘤發(fā)生[J];國(guó)外醫(yī)學(xué)(腫瘤學(xué)分冊(cè));1990年02期
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