本文選題：食管癌 + 自身抗原　； 參考：《河南中醫(yī)藥大學(xué)》2016年碩士論文

【摘要】：目的:本研究分析食管癌相關(guān)自身抗原CK13、CK16、CaD、ACTG2在食管癌四種證候中的表達(dá)分布規(guī)律,進(jìn)而探討清熱化痰方及其清熱和化痰兩個拆方對食管癌細(xì)胞增殖及其相關(guān)自身抗原表達(dá)的調(diào)節(jié)作用,為食管癌證候的分子病機(jī)和臨床用藥提供依據(jù)。方法:1.用食管癌自身抗原抗體和病人自身血清對食管癌組織進(jìn)行western blot分析比較;2.運(yùn)用免疫組化法分析CK13、CK16、CaD、ACTG2在四種證候食管癌正常組織、癌旁組織和癌組織中的表達(dá);3.運(yùn)用MTT法檢測清熱化痰方及其清熱和化痰兩個拆方對食管癌細(xì)胞EC109、EC9706和TE-1細(xì)胞增殖活性的影響;4.運(yùn)用流式細(xì)胞術(shù)分析清熱化痰方及其清熱和化痰兩個拆方對食管癌細(xì)胞EC109、EC9706和TE-1細(xì)胞周期的影響;5.運(yùn)用western blot分析清熱化痰方及其清熱和化痰兩個拆方對食管癌細(xì)胞EC109、EC9706和TE-1 CK13、CK16、CaD、ACTG2表達(dá)的影響。結(jié)果:1.痰氣交阻證、瘀血內(nèi)結(jié)證、津虧熱結(jié)證和氣虛陽微證食管癌病人的發(fā)病年齡不同,痰氣交阻證病人發(fā)病年齡為60.6±8.1歲,瘀血內(nèi)結(jié)證病人發(fā)病年齡為61.5±11.6歲,津虧熱結(jié)證病人發(fā)病年齡為65.6±8.1歲,氣虛陽微證病人發(fā)病年齡為68.4±7.3歲。2.痰氣交阻證、瘀血內(nèi)結(jié)證、津虧熱結(jié)證和氣虛陽微證組織中分別有自身抗原CK13、CaD、ACTG2和CK16的表達(dá),抗原自身抗體可以用于檢測食管癌組織自身抗原表達(dá);3.CaD在食管癌病人癌組織中表達(dá)陽性率和陽性表達(dá)水平高于正常組織和癌旁組織,其癌旁組織中表達(dá)陽性率和陽性表達(dá)水平高于正常組織,CaD在食管癌病人正常組織、癌旁組織、癌組織中表達(dá)陽性率分別為50.9%、62.2%、72.8%,陽性表達(dá)水平分別為0.0085±0.0048、0.0107±0.0056、0.0177±0.0103;CK16、ACTG2在癌組織中的陽性表達(dá)水平高于正常組織和癌旁組織,癌旁組織中陽性表達(dá)水平高于正常組織,CK16在食管癌病人正常組織、癌旁組織、癌組織中陽性表達(dá)水平分別為0.0076±0.0033、0.0158±0.0065、0.0356±0.0165,ACTG2在食管癌病人正常組織、癌旁組織、癌組織中陽性表達(dá)水平分別為0.0091±0.0039、0.0136±0.0043、0.0214±0.0110;CK13在癌組織中的陽性表達(dá)水平低于正常組織和癌旁組織,癌旁組織中陽性表達(dá)表達(dá)水平低于正常組織,CK13在食管癌病人正常組織、癌旁組織、癌組織中陽性表達(dá)水平分別為0.2053±0.0311、0.1633±0.0280、0.0412±0.0239;4.食管癌病人癌組織CK13、CK16、CaD、ACTG2的表達(dá)在四種證候之間存在差異:瘀血內(nèi)結(jié)證CaD陽性率為100%(6/6),其陽性表達(dá)水平為0.0364±0.0051,高于痰氣交阻證(29/34,0.0168±0.0088)、津虧熱結(jié)證(5/8,0.0159±0.0045)和氣虛陽微證(5/7,0.0166±0.0089);津虧熱結(jié)證ACTG2陽性表達(dá)水平為0.0346±0.0100,高于痰氣交阻證0.0201±0.0106、瘀血內(nèi)結(jié)證0.0244±0.0043和氣虛陽微證0.0235±0.0077;氣虛陽微證CK16陽性表達(dá)水平為0.0595±0.0131,明顯高于痰氣交阻證0.0325±0.0167、瘀血內(nèi)結(jié)證0.0227±0.0154、津虧熱結(jié)證0.0393±0.0068;氣虛陽微證CK13陽性表達(dá)水平為0.0174±0.0058,低于痰氣交阻證0.0452±0.0292、瘀血內(nèi)結(jié)證0.0385±0.0209和津虧熱結(jié)證0.0330±0.0251;CK13、CK16、CaD、ACTG2在食管癌病人正常組織和癌旁組織中表達(dá)的證候之間無差異;5.清熱化痰方及其清熱和化痰兩個拆方對食管癌細(xì)胞EC109、EC9706和TE-1的增殖均存在抑制作用,抑制作用從強(qiáng)到弱依次為全方組、清熱組、化痰組;6.清熱化痰方及其清熱和化痰兩個拆方對食管癌細(xì)胞EC109、EC9706和TE-1細(xì)胞周期均有影響:與空白對照組比較,EC109、TE-1細(xì)胞的清熱組、化痰組和全方組S期細(xì)胞均減少,其中化痰組變化最明顯;EC9706細(xì)胞清熱組S期細(xì)胞增加明顯,G2期細(xì)胞減少明顯,化痰組G1期細(xì)胞減少明顯,G2期細(xì)胞增加明顯,全方組G1期細(xì)胞增加明顯,G2期細(xì)胞減少明顯。7.清熱化痰方及其清熱和化痰兩個拆方可以不同程度地調(diào)節(jié)食管癌細(xì)胞EC109、EC9706和TE-1中CK13、CK16、CaD、ACTG2表達(dá),用藥組CK13表達(dá)高于空白組,用藥組CK16、CaD、ACTG2表達(dá)低于空白組。結(jié)論:1.食管癌病人癌組織均表達(dá)有自身抗原CK13、CK16、CaD、ACTG2,其中CK16、CaD、ACTG2在食管癌病變過程中表達(dá)上升,CK13表達(dá)下降;2.在食管癌不同證候病變過程中CK13、CK16、CaD、ACTG2均有改變,其中痰氣交阻證CK13表達(dá)改變突出,瘀血內(nèi)結(jié)證CaD表達(dá)改變突出,津虧熱結(jié)證ACTG2表達(dá)改變突出,氣虛陽微證CK16表達(dá)改變突出;3.清熱化痰方及其清熱和化痰兩個拆方對食管癌細(xì)胞Eca109、EC9706、TE-1增殖的抑制作用存在劑量依賴和時間依賴關(guān)系;4.清熱化痰方及其清熱和化痰兩個拆方對食管癌細(xì)胞Eca109、EC9706、TE-1的細(xì)胞周期有影響,其中Eca109、TE-1細(xì)胞各藥物組主要作用于S期,EC9706主要作用于S期和G2期;5.清熱化痰方及其清熱和化痰兩個拆方抑制食管癌細(xì)胞Eca109、EC9706和TE-1增殖與調(diào)節(jié)CK13、CK16、CaD、ACTG2蛋白表達(dá)相關(guān)為清熱化痰方治療食管癌提供了分子依據(jù);6.食管癌痰阻、瘀結(jié)、津虧、氣虛病機(jī)與自身抗原CK13、CK16、CaD、ACTG2表達(dá)改變相關(guān)。
[Abstract]:Objective: to analyze the expression and distribution of CK13, CK16, CaD and ACTG2 in the four syndromes of esophageal cancer, and to explore the regulation of the two dismantling prescriptions of clearing heat and phlegm and its heat clearing and resolving phlegm on the proliferation of esophageal cancer cells and the expression of their own antigen, for the molecular pathogenesis and clinical medication of the esophageal cancer syndrome. Methods: 1. Western blot analysis of esophageal cancer tissues was compared with the autoantigen antibody of esophageal cancer and the patient's own serum; 2. the expression of CK13, CK16, CaD, ACTG2 in four kinds of normal tissues of esophageal carcinoma, the expression of the para cancerous tissue and cancer tissues were analyzed by immunohistochemistry; 3. the MTT method was used to detect the heat clearing and eliminating phlegm, and the clearing heat and phlegm of the clearing heat and phlegm were detected by MTT method. The effects of two dismantling sides on the proliferation of EC109, EC9706 and TE-1 cells in esophageal cancer cells; 4. the effects of two dismantling sides of clearing heat and eliminating phlegm and clearing heat and phlegm on the cycle of EC109, EC9706 and TE-1 cells in esophageal cancer cells were analyzed by flow cytometry; 5. using Western blot to analyze the two dismantling sides of clearing heat and eliminating phlegm and eliminating phlegm and clearing heat and eliminating phlegm. The effect of EC109, EC9706 and TE-1 CK13, CK16, CaD, ACTG2 expression. Results: 1. sputum obstruction syndrome, blood stasis syndrome, Qi deficiency syndrome and Qi deficiency syndrome of esophageal cancer patients are different, the age of the patients with sputum resistance syndrome is 60.6 + 8.1 years old, the age of the patients with blood stasis syndrome is 61.5 + 11.6 years, and the patients with Tianjin deficiency heat syndrome Age is 65.6 + 8.1 years old, the age of qi deficiency syndrome patients is 68.4 + 7.3 years old.2. sputum resistance syndrome, blood stasis syndrome, Zimi heat syndrome and Qi deficiency syndrome, there are self antigens CK13, CaD, ACTG2 and CK16 expression, the antigen autoantibody can be used to detect the expression of self antigen of esophageal cancer tissue; 3.CaD in the cancer group of esophageal cancer patients The positive rate and positive expression level in the tissue were higher than that of the normal tissue and para cancerous tissue. The positive rate and positive expression level in the paracancerous tissues were higher than those of the normal tissue. The positive rates of CaD in normal tissues, para cancer tissues and cancer tissues of the patients with esophageal cancer were 50.9%, 62.2%, 72.8%, respectively, and the positive expression level was 0.0085 + 0.0048,0.0107, respectively. The positive expression level of ACTG2 in the cancer tissues was higher than that of normal tissue and para cancerous tissue. The positive expression level of the paracancerous tissues was higher than that of the normal tissue. The positive expression level of CK16 was 0.0076 + 0.0033,0.0158 + 0.0065,0.0356 + 0.0165 in the normal tissue, para cancer tissue and cancer tissue of the patients with esophageal cancer, and ACTG2 was in the diet, respectively. The positive expression level of ACTG2 in the cancer tissue was higher than that in the normal tissue. The positive expression level was 0.0091 + 0.0039,0.0136 + 0.0043,0.0214 + 0.0110 in normal tissue, para cancer tissue and cancer tissue in cancer patients. The positive expression level of CK13 in cancer tissues was lower than that of normal tissue and para cancer tissue. The positive expression level in para cancerous tissues was lower than that of normal tissue. CK13 was in normal tissue of the patients with esophageal cancer and in the para cancer group. The positive expression level in the carcinoma tissue was 0.2053 + 0.0311,0.1633 + 0.0280,0.0412 + 0.0239 respectively. The expression of CK13, CK16, CaD and ACTG2 in the carcinoma tissues of 4. patients with esophageal cancer was different between the four syndromes: the positive rate of CaD in the blood stasis syndrome was 100% (6/6), and the positive expression level was 0.0364 + 0.0051, which was higher than that of the sputum resistance syndrome (29/34,0.0168 + 0.0088). The syndrome of Jin deficiency heat syndrome (5/8,0.0159 + 0.0045) and Qi deficiency Yang Micro syndrome (5/7,0.0166 + 0.0089); the positive expression level of ACTG2 in Tianjin deficiency heat syndrome was 0.0346 + 0.0100, higher than that of sputum obstruction syndrome (0.0201 + 0.0106, 0.0244 + 0.0043 and Qi deficiency Yang Micro syndrome 0.0235 + 0.0077), and the positive expression level of qi deficiency Yang microsyndrome was 0.0595 + 0.0131, obviously higher than phlegm. The syndrome of gas crossing was 0.0325 + 0.0167, the syndrome in blood stasis was 0.0227 + 0.0154, the syndrome of Jin deficiency fever was 0.0393 + 0.0068, the positive expression level of CK13 in Qi deficiency syndrome was 0.0174 + 0.0058, lower than that of the sputum cross resistance syndrome, 0.0452 + 0.0292, the syndrome in stasis syndrome 0.0385 + 0.0209 and Jin deficiency syndrome 0.0330 + 0.0251, and CK13, CK16, CaD, ACTG2 in the normal tissue and para cancer of the patients with esophageal cancer. There were no differences between the syndromes expressed in the tissues; 5. the two dismantling sides of clearing heat and eliminating phlegm and clearing heat and resolving phlegm had inhibitory effect on the proliferation of EC109, EC9706 and TE-1 of esophageal cancer cells. The inhibitory effect was from strong to weak to the whole group, the clearing heat group and the phlegm group; 6. clearing heat and Phlegm Recipe and its clearing heat and phlegm two dismantling prescription on the esophageal cancer cells EC109, EC97 The cell cycle of 06 and TE-1 had an effect: compared with the blank control group, the cells in the EC109 and TE-1 cells were reduced in the clearing heat group, the phlegm group and the whole group were reduced in the S stage, of which the phlegm group had the most obvious change, the S phase cells in the EC9706 cell heat clearing group increased obviously, the G2 phase cells decreased obviously, the cell decrease in the phlegm group G1 phase decreased obviously, the G2 phase cells increased obviously, the G1 period of the whole group G1 period was more obvious. The cell increase obviously, G2 stage cell decrease obviously.7. clearing heat and phlegm prescription and two dismantling sides of clearing heat and phlegm can adjust esophageal cancer cells EC109, EC9706 and TE-1 CK13, CK16, CaD, ACTG2 expression, the expression of CK13 in the drug group is higher than that of the blank group, the drug group CK16, CaD, is lower than the blank group. Conclusion: all of the cancer tissues of the patients with esophageal cancer are all cancer tissue The expression of its own antigen CK13, CK16, CaD, ACTG2, CK16, CaD, ACTG2 in the process of esophageal cancer, the expression of CK13 decreased; 2. in the process of esophageal cancer, CK13, CK16, CaD, ACTG2 all changed. The change of CK16 expression was prominent. 3. the two dismantling prescriptions of clearing heat and eliminating phlegm and clearing heat and resolving phlegm had dose dependence and time dependence on the proliferation of Eca109, EC9706, and TE-1 in esophageal cancer cells; 4. the cell cycle of Eca109, EC9706 and TE-1 in the carcinoma cells of the esophagus and its clearing heat and the phlegm and its clearing heat and the resolving phlegm. Among them, Eca109, TE-1 cells were mainly used in S phase, EC9706 mainly acted on S and G2 phase; 5. the two dismantling sides of clearing heat and eliminating phlegm and clearing heat and resolving phlegm to inhibit the Eca109, EC9706 and TE-1 proliferation and regulation CK13, CK16, CaD, and CaD, and the expression phase for the treatment of esophageal cancer with molecular basis; 6. The pathogenesis of esophageal cancer phlegm blockage, stasis, deficiency of Qi and deficiency of Qi is related to the expression of CK13, CK16, CaD and ACTG2.
【學(xué)位授予單位】：河南中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R273

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