CAJ | 학술논문

目的观察蛋白激酶CK2抑制剂对人类非小细胞肺癌细胞活力及迁移能力的影响及对表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs)埃克替尼(icotinib)的靶向增敏作用.方法通过Western blot法检测蛋白激酶CK2各亚基在不同类型肺癌细胞株中的表达状态;免疫组织化学法检测CK2功能亚基在不同类型肺癌标本中得表达;采用噻唑蓝(MTT)法检测不同浓度梯度蛋白激酶CK2抑制剂醌茜素(Quinalizarin)对A549及H-460细胞活力的影响;同时用Transwell迁移实验检测其对这两种细胞迁移能力的影响;通过MTT法检测醌茜素与埃克替尼联合应用对不同EGFR表达状态的非小细胞肺癌细胞活力的影响.结果通过检测蛋白激酶CK2在不同类型肺癌细胞株及肺癌标本中的表达,发现该激酶广泛表达于各类型肺癌中.使用0、12.5、25.0、50.0 μmol/L终质量浓度的醌茜素抑制激酶活性后,MTT实验检测发现,随浓度增高A549及H460细胞活力均有明显抑制(P＜0.01);迁移实验中,随浓度增高A549细胞穿过人工基底膜的细胞数分别为(1041.0±115.9)、(705.3±74.7)、(451.3±55.0)、(128.3±38.0)个,H460穿过人工基底膜的细胞数分别为(924.0±65.6)、(593.7±69.2)、(954.7±24.7)、(193.0±27.2)个,均明显减少(P＜0.01).埃克替尼与醌茜素联合应用分别处理A549、H1650、H1975、PC9细胞,与单独使用埃克替尼比较,在50.0、100.0 μmol/L浓度时,各细胞株细胞活力明显下降(P＜0.01).结论蛋白激酶CK2在各种类型肺癌中广泛表达,通过降低CK2的激酶活性可明显抑制非小细胞肺癌细胞活力及迁移能力,并可增强埃克替尼对不同EGFR表达状态的非小细胞肺癌细胞的抑制作用.
목적 관찰단백격매CK2억제제대인류비소세포폐암세포활력급천이능력적영향급대표피생장인자수체락안산격매억제제(EGFR-TKIs)애극체니(icotinib)적파향증민작용.방법 통과Western blot법검측단백격매CK2각아기재불동류형폐암세포주중적표체상태;면역조직화학법검측CK2공능아기재불동류형폐암표본중득표체;채용새서람(MTT)법검측불동농도제도단백격매CK2억제제곤천소(Quinalizarin)대A549급H-460세포활력적영향;동시용Transwell천이실험검측기대저량충세포천이능력적영향;통과MTT법검측곤천소여애극체니연합응용대불동EGFR표체상태적비소세포폐암세포활력적영향.결과 통과검측단백격매CK2재불동류형폐암세포주급폐암표본중적표체,발현해격매엄범표체우각류형폐암중.사용0、12.5、25.0、50.0 μmol/L종질량농도적곤천소억제격매활성후,MTT실험검측발현,수농도증고A549급H460세포활력균유명현억제(P＜0.01);천이실험중,수농도증고A549세포천과인공기저막적세포수분별위(1041.0±115.9)、(705.3±74.7)、(451.3±55.0)、(128.3±38.0)개,H460천과인공기저막적세포수분별위(924.0±65.6)、(593.7±69.2)、(954.7±24.7)、(193.0±27.2)개,균명현감소(P＜0.01).애극체니여곤천소연합응용분별처리A549、H1650、H1975、PC9세포,여단독사용애극체니비교,재50.0、100.0 μmol/L농도시,각세포주세포활력명현하강(P＜0.01).결론 단백격매CK2재각충류형폐암중엄범표체,통과강저CK2적격매활성가명현억제비소세포폐암세포활력급천이능력,병가증강애극체니대불동EGFR표체상태적비소세포폐암세포적억제작용.
Objective To explore the effect of the protein kinase CK2 inhibitor on cell viability and migration of human non-small lung cancer cells and its enhanced role in sensitivity to Icotinib,one of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs).Methods The protein expression of individual CK2 subunits in different lung cancer cells was measured by Western blotting.The protein expression of the catalytic subunit-CK2α in different types of lung cancer tissues was detected by immunohistochemistry.Methyl thiazol tetrazolium (MTT) assays and Transwell migration assays were applied to assess the effect of CK2 inhibitor Quinalizarin on cell viability and migration capabilities of A549 and H460 respectively.The effect of the combination of Icotinib and Quinalizarin on cell viability of non-small lung cancer cell lines in the presence or absence of EGFR mutation was examined by MTT assays.Results Western bolting and immunohistochemistry revealed that protein kinase CK2 was overexpressed in different types of lung cancer cells and tissues.Quinalizarin,a CK2 inhibitor,could inhibit the cell viability of A549 and H460 cells in a dose-dependent manner within the range between 12.5 and 50.0 μmol/L (P ＜ 0.01).Along with the increased concentration of Quinalizarin,the migration of A549 and H460 was significantly suppressed (1 041.0 ± 115.9,705.3 ± 74.7,451.3 ± 55.0,128.3 ± 38.0 and 924.0 ± 65.6,593.7 ± 69.2,954.7 ± 24.7,193.0 ± 27.2,P ＜ 0.01).The combination of Icotinib and Quinalizarin could inhibit the cell viability of A549,H1650,H1975 and PC9 as compared with Icotinib used alone in the concentrations of 50 and 100 μmol/L (P ＜ 0.01).Conclusion Protein kinase CK2 was overexpressed in different types of lung cancer.Quinalizarin,one of the CK2 inhibitors can suppress the cell viability and migration capabilities of different types of lung cancer cells.It can also enhance the effect of Icotinib on cell viability of non-small lung cancer cell lines with or without EGFR mutation.