本文關(guān)鍵詞： 紫草素 縫隙連接蛋白32(Cx32) 細(xì)胞縫隙連接(GJ) 順鉑　出處：《新疆醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:在轉(zhuǎn)染并穩(wěn)定表達(dá)縫隙連接蛋白Cx32的宮頸癌Hela細(xì)胞上,觀察紫草素對縫隙連接GJ功能和縫隙連接蛋白Cx32的表達(dá)水平的影響,以及對順鉑細(xì)胞毒性作用的影響。方法:采用實(shí)時(shí)熒光定量PCR法和蛋白免疫印跡法對轉(zhuǎn)染并穩(wěn)定表達(dá)Cx32的Hela細(xì)胞系進(jìn)行驗(yàn)證;CCK-8法測定用于研究GJ功能作用的紫草素濃度;用細(xì)胞接種熒光示蹤法(Parachute assay)觀察不同濃度紫草素對GJ功能的影響;用蛋白免疫印跡(Western-blot)法研究紫草素對縫隙連接蛋白Cx32的蛋白表達(dá)的影響。用標(biāo)準(zhǔn)集落形成法(standard colony-forming assay)測定紫草素對順鉑細(xì)胞毒性的影響。結(jié)果:用多西環(huán)素培養(yǎng)的細(xì)胞有Cx32擴(kuò)增,也有Cx32的表達(dá),沒有用多西環(huán)素的細(xì)胞沒有Cx32擴(kuò)增,也沒有Cx32的表達(dá);對于不用多西環(huán)素誘導(dǎo)的細(xì)胞,即無Cx表達(dá)時(shí),紫草素4μM濃度以下對Hela細(xì)胞無毒性作用;用多西環(huán)素誘導(dǎo)的細(xì)胞,紫草素3μM濃度以下對Hela細(xì)胞無毒性作用;紫草素處理組細(xì)胞間熒光傳遞量在1-3μM隨濃度的增加而增加;紫草素在(1-3μM)能濃度依賴性增強(qiáng)GJ功能及Cx32的表達(dá)。在有GJ形成時(shí),3μM紫草素可以增強(qiáng)順鉑降低細(xì)胞集落形成率的作用。結(jié)論:驗(yàn)證Cx32質(zhì)粒轉(zhuǎn)染成功,構(gòu)建了可調(diào)控表達(dá)Cx32的細(xì)胞系。紫草素能增強(qiáng)Cx32組成的GJ功能。紫草素對GJ功能的增強(qiáng)作用與Cx32的表達(dá)有關(guān)。在有GJ形成的Hela細(xì)胞(生長融合),3μM紫草素可以增強(qiáng)順鉑的細(xì)胞毒性作用,有增敏的作用。在有GJ形成的Hela細(xì)胞(生長融合),紫草素可以增強(qiáng)順鉑的細(xì)胞毒性作用,具有增敏效果。
[Abstract]:Objective: to observe the effect of porphyrin on the function of gap junction GJ and the expression of gap junction protein Cx32 in cervical cancer Hela cells transfected and stably expressed gap junction protein (Cx32). Methods: the Hela cell lines transfected and stably expressed Cx32 were confirmed by real-time fluorescence quantitative PCR and Western blot. CCK-8 method was used to determine the concentration of porphyrin used to study the function of GJ. The effects of different concentrations of porphyrin on the function of GJ were observed by fluorescence tracer method. The effect of porphyrin on the expression of gap junction protein Cx32 was studied by Western blot assay. Standard colony-forming assay was used to determine the toxicity of porphyrin to cisplatin cells. Results: the cells cultured with doxycycline were amplified by Cx32. There was also Cx32 expression and no Cx32 amplification or Cx32 expression in cells without doxycycline; For the cells without doxycycline, that is, without CX expression, the concentration below 4 渭 M of porphyrin had no toxic effect on Hela cells. The cells induced by doxycycline had no toxic effect on Hela cells under the concentration of 3 渭 M of porphyrin. The fluorescence transfer between cells increased with the increase of concentration at 1-3 渭 M in the porphyrin treated group. Porphyrin could enhance the function of GJ and the expression of Cx32 in a concentration-dependent manner. 3 渭 M porphyrin could enhance the effect of cisplatin on cell colony formation. Conclusion: Cx32 plasmid transfection was successful. A cell line was constructed to regulate the expression of Cx32. Porphyrin can enhance the GJ function of Cx32. The enhancement of Rhizoctonin on GJ function is related to the expression of Cx32. Cells (. Growth and fusion). 3 渭 M porphyrin could enhance the cytotoxicity of cisplatin and enhance the sensitivity of cisplatin. In Hela cells formed with GJ (growth fusion), porphyrin could enhance the cytotoxicity of cisplatin. It has the effect of sensitizing.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張乃菊;余美玲;張鑫宇;祝曉光;;賴氨匹林增強(qiáng)Cx32/Cx26組成的Hela細(xì)胞縫隙連接功能[J];中國藥事;2014年07期

2 周海;伍春蓮;徐佳;彭聰;;紫草素對人結(jié)腸癌細(xì)胞Lovo增殖與凋亡的影響[J];西華師范大學(xué)學(xué)報(bào)(自然科學(xué)版);2014年02期

3 范高福;童旭輝;董淑英;蔣國君;陶亮;;鞣花酸對轉(zhuǎn)染并穩(wěn)定表達(dá)Cx32的HeLa細(xì)胞縫隙連接功能的影響[J];蚌埠醫(yī)學(xué)院學(xué)報(bào);2014年05期

4 郭雪興;陳劍群;;紫草素誘導(dǎo)消化系統(tǒng)腫瘤細(xì)胞凋亡的研究進(jìn)展[J];醫(yī)學(xué)綜述;2013年24期

5 張永紅;于慧敏;聶英坤;;紫草素對類風(fēng)濕關(guān)節(jié)炎滑膜成纖維細(xì)胞COX-2表達(dá)的影響[J];哈爾濱醫(yī)科大學(xué)學(xué)報(bào);2012年03期

6 童旭輝;董淑英;蔣國君;范高福;;縫隙連接蛋白Cx26/Cx32對依托泊苷抗腫瘤作用的影響[J];南方醫(yī)科大學(xué)學(xué)報(bào);2012年03期

7 李倩;陳韓英;廉宜君;田春芳;王曉琴;張波;鄭秋生;;新疆軟紫草多糖對小鼠免疫功能的影響[J];中國醫(yī)院藥學(xué)雜志;2011年10期

8 李麗君;余美玲;張素枝;王琴;陶亮;;木犀草素增強(qiáng)Cx32/Cx26組成的細(xì)胞縫隙連接功能[J];中山大學(xué)學(xué)報(bào)(醫(yī)學(xué)科學(xué)版);2011年03期

9 李治建;周凡;斯拉甫·艾白;劉發(fā);;紫草藥理作用研究進(jìn)展[J];中國中醫(yī)藥信息雜志;2010年11期

10 艾慶波;劉玉璇;國大亮;;中藥紫草的研究進(jìn)展[J];齊魯藥事;2010年10期

相關(guān)博士學(xué)位論文 前2條

1 白光振;紫草素在急性肺損傷中保護(hù)作用與機(jī)制的研究[D];第四軍醫(yī)大學(xué);2013年

2 王力娜;紫草素對局灶性腦缺血再灌注損傷小鼠的腦保護(hù)作用及其相關(guān)機(jī)制研究[D];河北醫(yī)科大學(xué);2013年

，

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