阿霉素联合二甲双胍应用对人舌癌细胞SCC-15侵袭迁移能力影响的实验研究

发布时间：2018-02-16 20:20

本文关键词： 舌癌阿霉素二甲双胍侵袭迁移　出处：《广西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：背景:舌癌是口腔颌面部最常见的恶性肿瘤之一,多数为鳞状上皮细胞癌,恶性程度高,生长快,浸润性强,在发病早期即出现颈部淋巴结转移,且易复发,预后较差;严重影响人类的身体健康和生活质量。目前舌癌的主要治疗手段是以手术扩大切除病灶为主并辅助放化疗治疗的综合序列治疗,尽管如此,舌癌的转移及复发率仍然较高,成为了舌癌治疗的重点难点;阿霉素是临床常用的广谱抗肿瘤药物,也是临床上用于头颈部鳞状细胞癌化疗的首选药物之一,临床化疗应用中容易产生耐药且随着药物剂量的增加其不良反应的严重程度也随之增加,这些都限制了恶性肿瘤的化疗效果;近年来的观察及研究发现广泛应用于II型糖尿病治疗的药物二甲双胍具有抗肿瘤作用,其药物安全性高耐受性好,相关研究表明其与多种常用化疗药物联合使用既能降低化疗药物使用剂量又能达到更好的抗癌作用效果。目的:应用阿霉素联合二甲双胍处理体外培养的人舌癌细胞,观察阿霉素联合二甲双胍用药与其二者单独用药相比对人舌癌细胞SCC-15侵袭、迁移能力等方面的影响,并初步探讨其可能的机制,为指导舌癌患者术后化疗用药提供理论依据,以期提高舌癌患者术后生存率。方法:体外常规培养人舌癌SCC-15细胞,给予阿霉素、二甲双胍及阿霉素联合二甲双胍进行干预作为实验组,无药物处理作为对照组。采用MTT法检测对照组及各实验组细胞存活率的影响;平板克隆形成实验检测对照组及各实验组细胞的体外克隆形成能力;Transwell侵袭小室实验观察对照组及各实验组细胞的体外侵袭能力;划痕-愈合实验观察对照组及各实验组细胞的体外迁移能力;q RT-PCR检测并比较对照组与各实验组细胞内金属基质蛋白酶MMP-2、MMP-9 m RNA的表达水平。结果:1.人舌癌细胞SCC-15的存活率随着阿霉素或二甲双胍浓度的递增而逐渐下降,与对照组相比,不同浓度阿霉素处理组24、48、72h的F值分别97.641、115.999、128.524,P0.01;不同浓度二甲双胍处理组24、48、72h的F值分别152.638、293.524、279.923,P0.01;与阿霉素或者二甲双胍单独用药相比,二者联合应用则更为显著的降低SCC-15细胞的存活率,差异有统计学意义(t值分别为-10.892和-6.311,P0.01)。2.阿霉素、二甲双胍均能抑制舌癌细胞SCC-15的体外克隆形成能力,与对照组相比,t值分别为-6.242和-14.519,P0.05;与阿霉素或二甲双胍单独用药相比,二者联合应用则更显著抑制SCC-15细胞的体外克隆形成能力,t值分别为-21.354和-13.610,P0.01。3.与对照组相比,在体外条件下0.05mg/L ADM能促进人舌癌SCC-15细胞的侵袭和迁移能力,t值分别为3.395和4.881,P0.05;二甲双胍(5mmol/L MET)则能抑制人舌癌SCC-15细胞的侵袭和迁移能力,t值分别为-4.593和-7.273,P0.05;与阿霉素、二甲双胍单独用药组相比两药联合应用(0.05mg/L ADM+5mmol/L MET)更为显著的抑制人舌癌SCC-15细胞的侵袭和迁移能力t值分别为-18.908、-23.655和-5.191、-6.592,P0.01。4.与对照组相比,阿霉素处理组细胞内MMP-2、MMP-9 m RNA的相对表达量增加,t值分别为32.659、43.246,P0.01;二甲双胍处理组细胞内MMP-2、MMP-9 m RNA的相对表达量则明显降低,t值分别为-23.989、-32.217,P0.01;与阿霉素、二甲双胍单独用药组相比两药联合应用处理组细胞内MMP-2、MMP-9 m RNA的相对表达量显著降低,t值分别为-33.352、-6.669和-67.214、-29.973,P0.01。结论:1.阿霉素和二甲双胍均能抑制SCC-15细胞的生长,并在一定浓度范围内呈时间-浓度依赖性,两药联合应用效果更为显著。2.二甲双胍能抑制人舌癌SCC-15细胞的侵袭及迁移,与阿霉素联合应用抑制效果更为显著;其主要机制可能是药物作用影响了相关基因的表达。
[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.

【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.86

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