本文關(guān)鍵詞： 舌癌 阿霉素 二甲雙胍 侵襲 遷移　出處：《廣西醫(yī)科大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：背景:舌癌是口腔頜面部最常見(jiàn)的惡性腫瘤之一,多數(shù)為鱗狀上皮細(xì)胞癌,惡性程度高,生長(zhǎng)快,浸潤(rùn)性強(qiáng),在發(fā)病早期即出現(xiàn)頸部淋巴結(jié)轉(zhuǎn)移,且易復(fù)發(fā),預(yù)后較差;嚴(yán)重影響人類(lèi)的身體健康和生活質(zhì)量。目前舌癌的主要治療手段是以手術(shù)擴(kuò)大切除病灶為主并輔助放化療治療的綜合序列治療,盡管如此,舌癌的轉(zhuǎn)移及復(fù)發(fā)率仍然較高,成為了舌癌治療的重點(diǎn)難點(diǎn);阿霉素是臨床常用的廣譜抗腫瘤藥物,也是臨床上用于頭頸部鱗狀細(xì)胞癌化療的首選藥物之一,臨床化療應(yīng)用中容易產(chǎn)生耐藥且隨著藥物劑量的增加其不良反應(yīng)的嚴(yán)重程度也隨之增加,這些都限制了惡性腫瘤的化療效果;近年來(lái)的觀察及研究發(fā)現(xiàn)廣泛應(yīng)用于II型糖尿病治療的藥物二甲雙胍具有抗腫瘤作用,其藥物安全性高耐受性好,相關(guān)研究表明其與多種常用化療藥物聯(lián)合使用既能降低化療藥物使用劑量又能達(dá)到更好的抗癌作用效果。目的:應(yīng)用阿霉素聯(lián)合二甲雙胍處理體外培養(yǎng)的人舌癌細(xì)胞,觀察阿霉素聯(lián)合二甲雙胍用藥與其二者單獨(dú)用藥相比對(duì)人舌癌細(xì)胞SCC-15侵襲、遷移能力等方面的影響,并初步探討其可能的機(jī)制,為指導(dǎo)舌癌患者術(shù)后化療用藥提供理論依據(jù),以期提高舌癌患者術(shù)后生存率。方法:體外常規(guī)培養(yǎng)人舌癌SCC-15細(xì)胞,給予阿霉素、二甲雙胍及阿霉素聯(lián)合二甲雙胍進(jìn)行干預(yù)作為實(shí)驗(yàn)組,無(wú)藥物處理作為對(duì)照組。采用MTT法檢測(cè)對(duì)照組及各實(shí)驗(yàn)組細(xì)胞存活率的影響;平板克隆形成實(shí)驗(yàn)檢測(cè)對(duì)照組及各實(shí)驗(yàn)組細(xì)胞的體外克隆形成能力;Transwell侵襲小室實(shí)驗(yàn)觀察對(duì)照組及各實(shí)驗(yàn)組細(xì)胞的體外侵襲能力;劃痕-愈合實(shí)驗(yàn)觀察對(duì)照組及各實(shí)驗(yàn)組細(xì)胞的體外遷移能力;q RT-PCR檢測(cè)并比較對(duì)照組與各實(shí)驗(yàn)組細(xì)胞內(nèi)金屬基質(zhì)蛋白酶MMP-2、MMP-9 m RNA的表達(dá)水平。結(jié)果:1.人舌癌細(xì)胞SCC-15的存活率隨著阿霉素或二甲雙胍濃度的遞增而逐漸下降,與對(duì)照組相比,不同濃度阿霉素處理組24、48、72h的F值分別97.641、115.999、128.524,P0.01;不同濃度二甲雙胍處理組24、48、72h的F值分別152.638、293.524、279.923,P0.01;與阿霉素或者二甲雙胍單獨(dú)用藥相比,二者聯(lián)合應(yīng)用則更為顯著的降低SCC-15細(xì)胞的存活率,差異有統(tǒng)計(jì)學(xué)意義(t值分別為-10.892和-6.311,P0.01)。2.阿霉素、二甲雙胍均能抑制舌癌細(xì)胞SCC-15的體外克隆形成能力,與對(duì)照組相比,t值分別為-6.242和-14.519,P0.05;與阿霉素或二甲雙胍單獨(dú)用藥相比,二者聯(lián)合應(yīng)用則更顯著抑制SCC-15細(xì)胞的體外克隆形成能力,t值分別為-21.354和-13.610,P0.01。3.與對(duì)照組相比,在體外條件下0.05mg/L ADM能促進(jìn)人舌癌SCC-15細(xì)胞的侵襲和遷移能力,t值分別為3.395和4.881,P0.05;二甲雙胍(5mmol/L MET)則能抑制人舌癌SCC-15細(xì)胞的侵襲和遷移能力,t值分別為-4.593和-7.273,P0.05;與阿霉素、二甲雙胍單獨(dú)用藥組相比兩藥聯(lián)合應(yīng)用(0.05mg/L ADM+5mmol/L MET)更為顯著的抑制人舌癌SCC-15細(xì)胞的侵襲和遷移能力t值分別為-18.908、-23.655和-5.191、-6.592,P0.01。4.與對(duì)照組相比,阿霉素處理組細(xì)胞內(nèi)MMP-2、MMP-9 m RNA的相對(duì)表達(dá)量增加,t值分別為32.659、43.246,P0.01;二甲雙胍處理組細(xì)胞內(nèi)MMP-2、MMP-9 m RNA的相對(duì)表達(dá)量則明顯降低,t值分別為-23.989、-32.217,P0.01;與阿霉素、二甲雙胍單獨(dú)用藥組相比兩藥聯(lián)合應(yīng)用處理組細(xì)胞內(nèi)MMP-2、MMP-9 m RNA的相對(duì)表達(dá)量顯著降低,t值分別為-33.352、-6.669和-67.214、-29.973,P0.01。結(jié)論:1.阿霉素和二甲雙胍均能抑制SCC-15細(xì)胞的生長(zhǎng),并在一定濃度范圍內(nèi)呈時(shí)間-濃度依賴性,兩藥聯(lián)合應(yīng)用效果更為顯著。2.二甲雙胍能抑制人舌癌SCC-15細(xì)胞的侵襲及遷移,與阿霉素聯(lián)合應(yīng)用抑制效果更為顯著;其主要機(jī)制可能是藥物作用影響了相關(guān)基因的表達(dá)。
[Abstract]:Background: tongue cancer is one of the most common malignant tumor in oral and maxillofacial region, the majority of squamous cell carcinoma, high malignancy, rapid growth, strong infiltration, i.e. cervical lymph node metastasis in the early onset and recurrence, the prognosis is poor; seriously affect human health and quality of life. The main treatment of tongue cancer the comprehensive treatment, operation sequence in the extended resection and adjuvant radiotherapy and chemotherapy in the treatment of the recurrence and metastasis of tongue cancer in spite of this, the rate is still high, has become a key and difficult treatment of tongue cancer; adriamycin is antitumor drugs commonly used in clinical spectrum, is one of the preferred drug for clinical chemotherapy of head and neck squamous cell carcinoma, multidrug resistance and with the severity of drug dose increase the adverse reaction increases prone to clinical chemotherapy, chemotherapy effect which limits malignant tumor in recent years; Observation and study to detect drug metformin is widely used in the treatment of type II diabetes has anti-tumor effect, the drug safety is well tolerated, and the related research shows that a variety of commonly used chemotherapy drugs combined with chemotherapy can reduce the dosage and can achieve better antitumor effect. Objective: human tongue cancer cell application adriamycin combined with metformin in vitro, observation of adriamycin combined with metformin monotherapy compared with the two drugs on the invasion of human tongue cancer cell line SCC-15, migration and other aspects, and explore its possible mechanism, to provide a theoretical basis for the chemotherapy medication guide tongue cancer patients after operation, in order to improve the survival rate of patients with tongue cancer after operation. Methods: in vitro cultured human tongue cancer SCC-15 cells to adriamycin, combined with metformin and adriamycin intervention as the experimental group, No drug treatment as the control group. MTT method was used to detect the effect of control group and survival rate of cells in each experimental group; clone formation assay in vitro cloning of the control group and the experimental group cells forming ability; Transwell invasion assay was used to observe the control group and the experimental group cells in vitro invasion; experimental observation and control group the experimental group cells in vitro scratch healing migration - Q; RT-PCR was detected and compared with control group MMP-2 in all groups of MMPs, the expression level of MMP-9 m RNA. Results: 1. SCC-15 cell survival rate with increasing concentrations of Adriamycin or metformin decreased compared with the control group, different the concentration of adriamycin treatment group 24,48,72h F-measure respectively 97.641115.999128.524, P0.01; different concentrations of metformin treatment group 24,48,72h F-measure respectively 152.638293.524279.923 and P0.01; Doxorubicin or metformin alone medication compared to the combination of the two methods is more significantly reduced the survival rate of SCC-15 cells, the difference was statistically significant (t = -10.892 and -6.311, P0.01).2. adriamycin, metformin can inhibit cancer cells SCC-15 in vitro clone forming ability, compared with the control group, t value respectively. -6.242 and -14.519, P0.05; compared with doxorubicin or metformin monotherapy, the combination of the two methods in vitro cloning is more significantly inhibited the formation ability of SCC-15 cells, t values were -21.354 and -13.610, P0.01.3. compared with the control group, can promote the invasion and migration of human tongue cancer SCC-15 cells in vitro under the condition of 0.05mg/L ADM, t were 3.395 and 4.881, P0.05 (5mmol/L MET); metformin can inhibit the invasion and migration of human tongue cancer SCC-15 cells, t values were -4.593 and -7.273, P0.05 and adriamycin, two a; The biguanide monotherapy group compared with the combination of two drugs (0.05mg/L ADM+5mmol/L MET) is more significant inhibition of human tongue cancer SCC-15 cell invasion and migration ability of T = -18.908, -23.655 and -5.191, -6.592, P0.01.4. compared with the control group, adriamycin treatment group MMP-2 cells, the relative expression of MMP-9 m increased the amount of RNA, t = 32.659,43.246, P0.01; metformin treatment group of MMP-2 cells, the relative expression of MMP-9 m RNA were significantly lower, t = -23.989, -32.217, P0.01; and adriamycin, metformin alone treatment group compared with two drug combination treatment group of intracellular MMP-2, relative expression of MMP-9 m significantly decreased RNA, t the values were -33.352, -6.669 and -67.214, -29.973, P0.01.. Conclusion: 1. adriamycin and metformin can inhibit the growth of SCC-15 cells in a time - and concentration dependent in a certain range of concentration, the combination of two drugs The effect is more significant..2. metformin can inhibit the invasion and migration of human tongue cancer SCC-15 cells, and combined with adriamycin, the inhibitory effect is more significant. The main mechanism may be that the action of drugs affects the expression of related genes.

【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R739.86

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本文編號(hào)：1516353