薏苡仁注射液联合X线照射对人肺腺癌细胞A549抑制作用的机制研究

Inhibiting Mechanism of Coix Lacryma-Jobi Injection Combined with Radiotherapy in Human Lung Carcinoma Cells A549

目的探讨薏苡仁注射液(康莱特)联合X线照射对人肺腺癌细胞A549抑制作用的机制。方法体外培养人肺腺癌细胞A549,分成4组:A549对照组(给予1640培养液);A549康莱特组(给予20μl康莱特);A549照射组(照射4Gy剂量的X线);A549联合组(给予20μl康莱特,同时4Gy的X线照射)。正常细胞系人脑微血管内皮细胞(HBMEC),分成2组:HBMEC对照组(给予0.9%氯化钠溶液);HBMEC康莱特组(给予20μl康莱特)。磺酰罗丹明(SRB)染色法检测康莱特及X线照射对各组细胞的抑制率;逆转录-聚合酶链反应(RT-PCR)法检测增殖、凋亡相关蛋白增殖细胞核抗原(PCNA)、p53、Bcl-2、BaxmRNA在各组细胞中表达情况;蛋白质印迹(West-ernblot)法检测蛋白表达水平。将各组结果进行比较。结果康莱特联合放疗对6组细胞体外抑制率、PCNA、p53、Bcl-2、BaxmRNA及4种因子的蛋白表达比较,差异均有统计学意义(P<0.05)。其中与A549对照组比较,A549康莱特组、A549照射组、A549联合组的抑制率、PCNA和Bcl-2mRNA及4种因子的蛋白表达降低,p53、BaxmRNA及其蛋白表达升高,差异均有统计学意义(P<0.05);HBMEC康莱特组与HBMEC对照组比较,抑制率、4种因子表达比较,差异均无统计学意义(P>0.05)。结论康莱特可通过调控肿瘤细胞的增殖凋亡来抑制肺腺癌细胞生长,该药与放射联合治疗可进一步增强对肿瘤的抑制作用,探讨康莱特作用机制对指导临床肺腺癌治疗有一定的价值。

Objective To investigate the inhibiting mechanism of Coix Lacryma-Jobi Injection（Kanglaite）combined with external irradiation in human lung carcinoma cells A549.Methods Lung carcinoma cells A549 were divided randomly into groups A549 control （given 1640 solution）,A549 kanglaite （given 20 μl kanglaite）,A549 irradiation （ given 4 Gy X ray ）,A549 combination （given 20 μl kanglaite and 4 Gy X ray）.Human brain microvascular endothelial cells （HBMEC） were divided randomly into groups HBMEC control （given 0.9% sodium chloride solution）,HBMEC kanglaite （given 20 μl kanglaite）.The inhibition rates were determined by SRB staining in vitro,and the expressions of proliferating cell nuclear antigen（PCNA）,p53,Bcl-2,Bax by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot.Results Kanglaite combined with irradiation had obvious influence on the inhibition rate,the expressions （mRNA and proteins） of PCNA,p53,Bcl-2,Bax （P0.05）.Compared with A549 control group,the inhibition rate and the expressions of PCNA,Bcl-2 decreased,the expressions of p53,Bax increased in groups A549 Kanglaite,A549 irradiation,A549 Kanglaite combination （P0.05）,and there was no significant difference between groups HBMEC Kanglaite and HBMEC control （P0.05）.Conclusion Kanglaite can inhibit the growth of A549 cells by regulating proliferation and apoptosis of tumor cells.Combination of Kanglaite and CDDP can increase chemosensitivity of A549 cells.