本文選題：重樓 + 胃癌��； 參考：《南京中醫(yī)藥大學(xué)》2017年博士論文

【摘要】：[目的]研究重樓乙醇提取物(Parispolyphlla ethanol extract,PPE)對ICR小鼠的急性毒性劑量,探討重樓在腫瘤綜合治療中的應(yīng)用,并考察其體外的抗腫瘤作用及機制;探討胃癌患者的術(shù)前血清腫瘤標(biāo)記物水平和手術(shù)后脈管內(nèi)癌栓之間的關(guān)系,初步探討胃癌臨床應(yīng)用中藥聯(lián)合氟尿嘧啶類化療的療效及其對療效相關(guān)分子調(diào)節(jié)作用。[方法]1、用ICR小鼠進行急性毒性實驗,以生存狀態(tài)、血液學(xué)指標(biāo)變化、腹腔臟器變化為觀察指標(biāo),并計算PPE的半數(shù)致死量(median leath death,LD50)劑量。2、用MTT法及細胞克隆形成方法檢測PPE,以及PPE聯(lián)合放療對人胃癌細胞MKN-45的生長抑制;用PI染色法檢測PPE,以及PPE聯(lián)合放療對人胃癌細胞株MKN-45細胞周期的影響;Western-blot檢測PPE對凋亡相關(guān)蛋白水平的影響;運用RT-PCR法檢測PPE對氟尿嘧啶類藥物敏感基因胸苷酸合成酶(thymidylatesynthase,TS)的mRNA水平的變化。3、通過對我院普通外科進行胃癌根治性手術(shù)的1005例患者,檢測手術(shù)前一周內(nèi)的外周血腫瘤標(biāo)志物CEA、CA19-9、CA242、CA72-4、CA125的水平,并對術(shù)后標(biāo)本進行包括脈管癌栓在內(nèi)的多項病理學(xué)檢查。入組患者基線水平數(shù)據(jù)按照各外周血腫瘤標(biāo)志物的狀況被分組,分析每個亞組標(biāo)志物水平對脈管癌栓狀態(tài)的影響。評價各亞組腫瘤標(biāo)志物水平預(yù)測脈管癌栓的敏感性、特異性、陽性預(yù)測價值、陰性預(yù)測價值;追蹤胃癌根治術(shù)Ⅲ期的輔助化療患者共270名,化療前后患者均進行外周血TS mRNA檢測。將入組患者分成中藥結(jié)合化療和單純化療組。入組患者每月檢測外周血腫瘤標(biāo)志物CEA、CA19-9、CA242、CA72-4、CA125的水平,每三月復(fù)查影像學(xué)評價病情進展。比較兩組患者的外周血腫瘤學(xué)標(biāo)志物變化情況,1年內(nèi)無復(fù)發(fā)生存率,以及化療前后外周血TS mRNA水平變化。[結(jié)果]1、單次給藥后獲得PPE對ICR小鼠的LD50值約為:12mg/ml,給藥劑量越接近LD50值的小鼠,其白細胞較正常小鼠組明顯下降,紅細胞及血小板無明顯變化,藥物對小鼠臟器未見明顯毒性作用。2、PPE對人胃癌細胞MKN-45的抑制率呈時間和劑量依賴性;重樓聯(lián)合放療可以降低腫瘤細胞的生存率;重樓醇提物單獨作用將MKN-45細胞阻滯在S期,而與放療聯(lián)合細胞周期阻滯在G2/M期;凋亡分析顯示PPE可以誘導(dǎo)腫瘤細胞凋亡,同時PPE與放療以協(xié)同作用方式誘導(dǎo)腫瘤細胞凋亡。PPE可下調(diào)Bcl-2蛋白的表達及上調(diào)Bax蛋白的表達。PPE可下調(diào)TS mRNA水平,在MKN-45細胞系中下調(diào)至0.59 ±0.07倍。3、入組的1005名患者中共有540名(53.73%)患者發(fā)現(xiàn)術(shù)后存在血管內(nèi)癌栓。對于至少有一項外周血腫瘤標(biāo)志物升高的患者來說,其脈管癌栓發(fā)生危險性是沒有腫瘤標(biāo)志物升高患者的2.9倍。對于至少有四項外周血腫瘤標(biāo)志物升高的患者,對比沒有腫瘤標(biāo)志物升高的患者,其脈管癌栓發(fā)生風(fēng)險是其16.4倍;輔助化療聯(lián)合中藥治療組與單純輔助化療組比較,6周期輔助化療后外周血腫瘤標(biāo)志物較化療前上升超過20%者分別有27名(19.3%)及34名(26.2%,P=0.178);1年內(nèi)病情復(fù)發(fā)的患者分別有31名(22.1%)及43名(33.1%,P=0.044);兩組間化療前外周血TS mRNA表達水平無顯著差異(聯(lián)合組:1.31,化療組:1.28,P=0.93),6周期化療后單純化療組患者外周血TS mRNA表達平均水平較化療前上升(1.28,1.42,P=0.43),化療聯(lián)合中藥治療組患者外周血TS mRNA表達平均水平較化療前下降(1.31,0.98,P=0.12),但均未達到統(tǒng)計學(xué)顯著性差異。[結(jié)論]1、PPE具有毒性,可能是通過血液白細胞的變化所致,對于腹腔臟器沒有明顯影響,臨床應(yīng)用需關(guān)注血液學(xué)檢測指標(biāo)。2、PPE具有顯著的體外抗腫瘤作用,其抗腫瘤作用的機理可能與影響細胞周期,下調(diào)Bcl-2蛋白的表達及上調(diào)Bax蛋白的表達來誘導(dǎo)腫瘤細胞凋亡,下調(diào)TS基因表達水平提高化療藥物敏感性等作用有關(guān);PPE與放療結(jié)合可以阻滯細胞周期,誘導(dǎo)腫瘤細胞凋亡,其機制可能與釋放活性氧相關(guān)。重樓是一味在腫瘤綜合治療中非常有前景的中藥。3、術(shù)前外周血腫瘤標(biāo)記物可以用于預(yù)測術(shù)后靜脈癌栓狀態(tài),對于術(shù)前血漿腫瘤標(biāo)志物升高的患者我們需要考慮更加積極的術(shù)前治療。4、胃癌Ⅲ期根治術(shù)后輔助化療患者中,聯(lián)合中藥治療較單純化療能顯著減少腫瘤復(fù)發(fā)風(fēng)險,其機制可能與中藥能夠降低患者TS mRNA表達從而提高氟尿嘧啶類化療藥物療效相關(guān)。
[Abstract]:[Objective] to study the acute toxic dose of Parispolyphlla ethanol extract (PPE) on ICR mice, to explore the application of heavy building in the comprehensive treatment of tumor, and to investigate the anti-tumor effect and mechanism in vitro, and to explore the relationship between the level of serum tumor markers before operation and the relationship between the tumor thrombus in the pulse tube after operation. The effect of the combination of traditional Chinese medicine and fluorouracil in the clinical application of gastric cancer and the regulating effect on the therapeutic effect of fluorouracil. [methods]1, ICR mice were used for acute toxicity test, the survival state, the change of Hematology Index, the change of abdominal organ as the observation index, and the dosage of median leath death, LD50), and the dosage of.2, and MTT. The method and cell clone formation method were used to detect the growth inhibition of PPE and PPE combined with radiotherapy for human gastric cancer cell MKN-45; the effect of PPE and PPE combined with radiotherapy on the MKN-45 cell cycle of human gastric cancer cell line were detected by PI; Western-blot was used to detect the effect of PPE on the level of apoptosis related proteins; PPE on fluorouracil was detected by RT-PCR method. Changes in the mRNA level of the thymidylatesynthase (TS) of the substance sensitive gene.3. The level of peripheral blood tumor markers, CEA, CA19-9, CA242, CA72-4, and CA125, were detected in 1005 patients undergoing radical gastrectomy for gastric cancer in general surgery in our hospital, and many of the postoperative specimens were included in the postoperative specimens including the vascular tumor thrombus. The baseline level data of the patients were grouped according to the status of the peripheral blood tumor markers, and the effects of each subgroup marker level on the vascular tumor thrombus status were analyzed. The sensitivity, specificity, positive predictive value and negative predictive value of the tumor markers in each subgroup were evaluated. 270 patients were treated with adjuvant chemotherapy in stage III, and peripheral blood TS mRNA detection was performed before and after chemotherapy. The patients were divided into groups of Chinese medicine combined with chemotherapy and chemotherapy alone. The patients enrolled in the group were detected the level of peripheral blood tumor markers CEA, CA19-9, CA242, CA72-4, CA125 every month, and evaluated the progress of the disease every March. Compare the two groups. There was no recurrence survival rate within 1 years, and the change of TS mRNA in peripheral blood before and after chemotherapy. [results]1, the LD50 value of PPE to ICR mice after a single dose of drug was about 12mg/ml, the more the dosage was closer to the LD50 value, the leukocytes were significantly lower than those of the normal mice, and the red blood cells and platelets had no obvious changes. The drug has no obvious toxic effect on mouse organs.2, and the inhibitory rate of PPE on human gastric cancer cell MKN-45 is time and dose dependent; heavy building combined with radiotherapy can reduce the survival rate of tumor cells; heavy building alcohol extracts separate MKN-45 cells in S phase alone, and the cell cycle block in G2/M phase with radiotherapy; apoptosis analysis shows PPE. The apoptosis of tumor cells could be induced, and the apoptosis of tumor cells was induced by PPE and radiotherapy in a synergistic manner..PPE could down regulate the expression of Bcl-2 protein and up regulate the expression of Bax protein,.PPE can down regulate the level of TS mRNA, down to 0.59 + 0.07 times.3 in MKN-45 cell line, and 540 (53.73%) patients in the 1005 patients of the group were found to exist after the operation. Intravascular thrombus. For patients with at least one increase in peripheral blood tumor markers, the risk of the vascular tumor thrombus was 2.9 times as high as those without a tumor marker. For patients with at least four peripheral blood tumor markers, the risk of the vascular tumor thrombus was 16. compared to those who had no higher tumor markers. 4 times, compared with the adjuvant chemotherapy group and the simple adjuvant chemotherapy group, there were 27 (19.3%) and 34 (26.2%, P=0.178) of peripheral blood tumor markers in the 6 cycle adjuvant chemotherapy than before chemotherapy, and 31 (22.1%) and 43 (33.1%, P=0.044) in 1 years, respectively, and TS mRNA of peripheral blood before chemotherapy before chemotherapy, and TS mRNA before chemotherapy. There was no significant difference in expression level (1.31, 1.31, chemotherapy group: 1.28, P=0.93). The average level of TS mRNA expression in peripheral blood of patients with chemotherapy after 6 cycles was higher than that before chemotherapy (1.28,1.42, P=0.43). The level of TS mRNA expression in peripheral blood of patients with chemotherapy combined with Chinese medicine treatment group was lower than that before chemotherapy (1.31,0.98, P=0.12), but no statistics were reached. [conclusion]1, PPE is toxic and may be caused by changes in blood leucocyte, it has no obvious influence on abdominal organs. Clinical application should pay attention to hematology detection index.2, PPE has significant antitumor effect in vitro, its anti-tumor mechanism can affect cell cycle, down regulation of Bcl-2 protein expression and The expression of Bax protein can induce apoptosis of tumor cells and reduce the level of TS gene expression to improve the chemosensitivity. The combination of PPE and radiotherapy can block cell cycle and induce apoptosis of tumor cells. The mechanism may be related to the release of reactive oxygen species. The anterior peripheral blood tumor markers can be used to predict the postoperative venous tumor thrombus status. For patients with elevated plasma tumor markers, we need to consider more active preoperative treatment of.4. In patients with adjuvant chemotherapy after stage III radical gastrectomy, the combination of traditional Chinese medicine and chemotherapy can significantly reduce the risk of tumor recurrence. The mechanism may be related to the possible mechanism. Chinese medicine can reduce the expression of TS mRNA in patients, thereby enhancing the efficacy of fluorouracil chemotherapy drugs.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R735.2

【參考文獻】

相關(guān)期刊論文 前10條

1 于思;曹治興;楊雨婷;白姣姣;彭成;李玉芝;;重樓皂苷Ⅰ誘導(dǎo)G_2/M期阻滯及干擾微管結(jié)構(gòu)抗結(jié)腸癌HCT116細胞作用機制[J];中國實驗方劑學(xué)雜志;2017年06期

2 Henrik Johansson;Roland Andersson;Monika Bauden;Sarah Hammes;Stefan Holdenrieder;Daniel Ansari;;Immune checkpoint therapy for pancreatic cancer[J];World Journal of Gastroenterology;2016年43期

3 宋延平;許建秦;張宇晨;王磊;趙林濤;;野生和人工培育陜重樓對荷瘤小鼠實體瘤及免疫因子的影響[J];陜西中醫(yī);2016年09期

4 孫家艷;茅淵;朱靜;;我院近兩年抗腫瘤中成藥及輔助用藥的應(yīng)用分析[J];現(xiàn)代中藥研究與實踐;2016年02期

5 張珂;鄧清華;馬勝林;;重樓醇提取物對胃癌SGC-7901細胞增殖和凋亡的影響[J];中華中醫(yī)藥學(xué)刊;2016年01期

6 趙聰;;中醫(yī)健脾祛瘀療法在胃癌的臨床治療分析[J];中醫(yī)臨床研究;2015年03期

7 江皓;趙鵬軍;馮建國;花永虹;馬勝林;;重樓皂苷Ⅰ對胰腺癌PANC-1細胞體外放射敏感性的影響[J];腫瘤學(xué)雜志;2014年06期

8 張元清;舒鵬;;運用健脾法治療胃癌研究進展[J];遼寧中醫(yī)藥大學(xué)學(xué)報;2014年02期

9 Zhi-Xue Zheng;Yu Sun;Zhao-De Bu;Lian-Hai Zhang;Zi-Yu Li;Ai-Wen Wu;Xiao-Jiang Wu;Xiao-Hong Wang;Xiao-Jing Cheng;Xiao-Fang Xing;Hong Du;Jia-Fu Ji;;Intestinal stem cell marker LGR5 expression during gastric carcinogenesis[J];World Journal of Gastroenterology;2013年46期

10 陳源紅;曾怡;覃艷春;莫頌軼;;重樓水提物對肝癌細胞HepG2增殖及凋亡的影響[J];右江民族醫(yī)學(xué)院學(xué)報;2013年05期

，

本文編號：2055426