结肠癌转移相关基因1表达与肺鳞状细胞癌和肺腺癌细胞顺铂耐药性的相关性

Relationship between the expression of metastasis-associated in colon cancer 1 and cisplatin resistance in pulmonary squamous cell carcinoma and pulmonary adenocarcinoma cells

目的探讨结肠癌转移相关基因1(MACC1)与肺鳞状细胞癌和肺腺癌细胞顺铂耐药性的关系。方法收集2018年1月至2019年12月郑州大学第五附属医院手术切除的新鲜肺腺癌组织标本37例和肺鳞状细胞癌组织标本23例,所有患者术前未行放射治疗和化学治疗。取新鲜癌组织行原代细胞体外培养。取第2代肺鳞状细胞癌和肺腺癌细胞,将癌细胞分别均分为顺铂组和对照组,顺铂组细胞以含顺铂的培养液继续培养72 h,对照组细胞按原代细胞培养液继续培养72 h,采用甲基噻唑基四唑(MTT)法检测肺鳞状细胞癌和肺腺癌细胞对顺铂的敏感性。取新鲜癌组织,采用免疫组织化学染色检测肺腺癌和肺鳞状细胞癌组织中MACC1蛋白的表达。取第2代肺鳞状细胞癌和肺腺癌细胞,反转录-聚合酶链反应法检测肺腺癌和肺鳞状细胞癌细胞中MACC1 mRNA的表达。结果37例肺腺癌患者对顺铂的敏感率为59.46%(22/37),耐药率为40.54%(15/37);23例肺鳞状细胞癌患者对顺铂的敏感率为52.17%(12/23),耐药率为47.83%(11/23);肺腺癌与肺鳞状细胞癌患者对顺铂的敏感性比较差异无统计学意义(χ2=2.849,P>0.05)。高、中、低分化肺腺癌对顺铂的耐药率分别为41.67%(5/12)、42.11%(8/19)、33.33%(2/6),高、中、低分化肺腺癌对顺铂的耐药率比较差异无统计学意义(χ2=2.916,P>0.05),高、中、低分化肺鳞状细胞癌对顺铂的耐药率分别为50.00%(2/4)、46.67%(7/15)、50.00%(2/4),高、中、低分化肺鳞状细胞癌对顺铂的耐药率比较差异无统计学意义(χ2=1.527,P>0.05);高、中、低分化的肺腺癌与肺鳞状细胞癌对顺铂的耐药率比较差异均无统计学意义(χ2=2.014、1.034、1.679,P>0.05)。Ⅰ、Ⅱ、Ⅲa、Ⅲb、Ⅳ期肺腺癌对顺铂的耐药率分别为0.00%(0/2)、33.33%(1/3)、36.36%(4/11)、38.46%(5/13)、62.50%(5/8);随着TNM分期升高,肺腺癌对顺铂的耐药率升高(χ2=21.417,P<0.05)。Ⅰ、Ⅱ、Ⅲa、Ⅲb、Ⅳ期肺鳞状细胞癌对顺铂的耐药率分别为0.00%(0/1)、33.33%(1/3)、44.44%(4/9)、50.00%(4/8)、100.00%(2/2);随着TNM分期升高,肺鳞状细胞癌对顺铂的耐药率升高(χ2=19.034,P<0.05)。Ⅰ、Ⅱ、Ⅲa期肺腺癌与肺鳞状细胞癌对顺铂的耐药率比较差异无统计学意义(χ2=2.327、0.159、0.827,P>0.05);Ⅲb、Ⅳ期肺腺癌对顺铂的耐药率显著低于肺鳞状细胞癌(χ2=8.361、10.357,P<0.05)。肺腺癌和肺鳞状细胞癌组织中MACC1蛋白表达比较差异无统计学意义(χ2=3.119,P>0.05)。肺腺癌和肺鳞状细胞癌细胞中MACC1 mRNA相对表达量分别为1.217±0.012、1.137±0.011,肺腺癌与肺鳞状细胞癌细胞中MACC1 mRNA相对表达量比较差异无统计学意义(t=12.137,P>0.05)。肺腺癌和肺鳞状细胞癌组织中MACC1蛋白表达与顺铂耐药性呈正相关(r=0.747、0.681,P<0.05)。结论肺腺癌和肺鳞状细胞癌对顺铂的耐药率较高,肺腺癌和肺鳞状细胞癌组织中MACC1蛋白表达与顺铂耐药性呈正相关,MACC1可能参与了肺腺癌和肺鳞状细胞癌对顺铂耐药的发生机制。

Objective To investigate the relationship between the expression of metastasis-associated in colon cancer 1(MACC1)and cisplatin resistance in pulmonary squamous cell carcinoma and pulmonary adenocarcinoma cells.Methods The fresh cancer tissue specimens of 37 cases of pulmonary adenocarcinoma and 23 cases of pulmonary squamous cell carcinoma were collected in the Fifth Affiliated Hospital of Zhengzhou University from January 2018 to December 2019.All patients did not receive radiotherapy and chemotherapy before operation.The fresh cancer tissues were taken for primary cell culture in vitro.The second generation cancer cells of pulmonary squamous cell carcinoma and pulmonary adenocarcinoma were collected and divided into cisplatin group and control group.The cells in the cisplatin group were cultured in medium containing cisplatin for 72 hours,while the cells in the control group were cultured in primary culture medium for 72 hours.The sensitivity of pulmonary squamous cell carcinoma and pulmonary adenocarcinoma cells to cisplatin was detected by methyl thiazolyl tetrazolium(MTT)method.The fresh cancer tissues was obtained,and the expression of MACC1 protein in pulmonary squamous cell carcinoma and pulmonary adenocarcinoma tissues was detected by immunohistochemistry.The second generation cancer cells of pulmonary squamous cell carcinoma and pulmonary adenocarcinoma were collected,and the expression of MACC1 mRNA in pulmonary squamous cell carcinoma and pulmonary adenocarcinoma cells was detected by reverse transcription-polymerase chain reaction.Results The sensitivity rate of 37 cases of pulmonary adenocarcinoma to cisplatin was 59.46%(22/37),and the drug resistance rate was 40.54%(15/37).The sensitivity rate of 23 cases of pulmonary squamous cell carcinoma to cisplatin was 52.17%(12/23),and the drug resistance rate was 47.83%(11/23).There was no significant difference in the sensitivity of pulmonary adenocarcinoma and pulmonary squamous cell carcinoma to cisplatin(χ2=2.849,P>0.05).The resistance rate of highly,moderately and poorly differentiated pulmonary adenocarcinoma to cisplatin was 41.67%(5/12),42.11%(8/19)and 33.33%(2/6),respectively.There was no significant difference in the resistance rate of highly,moderately and poorly differentiated pulmonary adenocarcinoma to cisplatin(χ2=2.916,P>0.05).The resistance rate of highly,moderately and poorly differentiated pulmonary squamous cell carcinoma to cisplatin was 50.00%(2/4),46.67%(7/15)and 50.00%(2/4),respectively.There was no significant difference in the resistance rate of highly,moderately and poorly differentiated pulmonary squamous cell carcinoma to cisplatin(χ2=1.527,P>0.05).There was no significant difference in cisplatin resistance rate between the highly,moderately and poorly differentiated pulmonary adenocarcinoma and pulmonary squamous cell carcinoma(χ2=2.014,1.034,1.679;P>0.05).The resistance rate of pulmonary adenocarcinoma of stageⅠ,Ⅱ,Ⅲa,Ⅲb andⅣto cisplatin was 0.00%(0/2),33.33%(1/3),36.36%(4/11),38.46%(5/13)and 62.50%(5/8),respectively.With the increase of TNM stage,the resistance rate of pulmonary adenocarcinoma to cisplatin was higher(χ2=21.417,P<0.05).The resistance rate of pulmonary squamous cell carcinoma of stageⅠ,Ⅱ,Ⅲa,Ⅲb andⅣto cisplatin was 0.00%(0/1),33.33%(1/3),44.44%(4/9),50.00%(4/8)and 100.00%(2/2),respectively.With the increase of TNM stage,the resistance rate of pulmonary squamous cell carcinoma to cisplatin was higher(χ2=19.034,P<0.05).There was no significant difference in cisplatin resistance rate between the pulmonary adenocarcinoma and pulmonary squamous cell carcinoma of stageⅠ,Ⅱ,Ⅲa(χ2=2.327,0.159,0.827;P>0.05).The cisplatin resistance rate of pulmonary adenocarcinoma of stageⅢb andⅣwas significantly lower than that of pulmonary squamous cell carcinoma(χ2=8.361,10.357;P<0.05).There was no significant difference in the expression of MACC1 protein between pulmonary adenocarcinoma and pulmonary squamous cell carcinoma(χ2=3.119,P>0.05).The relative expression of MACC1 mRNA in pulmonary adenocarcinoma and pulmonary squamous cell carcinoma cells was 1.217±0.012 and 1.137±0.011,respectively.There was no significant difference in the relative expression of MACC1 mRNA between pulmonary adenocarcinoma and pulmonary squamous cell carcinoma cells(t=12.137,P>0.05).The expression of MACC1 protein in pulmonary adenocarcinoma and pulmonary squamous cell carcinoma was positively correlated with cisplatin resistance(r=0.747,0.681;P<0.05).Conclusion The resistance rate of pulmonary adenocarcinoma and pulmonary squamous cell carcinoma to cisplatin is higher.The expression of MACC1 protein in pulmonary adenocarcinoma and pulmonary squamous cell carcinoma is positively correlated with cisplatin resistance.MACC1 may be involved in the resistance mechanism of pulmonary adenocarcinoma and pulmonary squamous cell carcinoma to cisplatin.