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目的探讨脾源性酪氨酸激酶(Syk)基因的再表达对裸鼠乳腺癌肿瘤生长和转移的影响。方法5氮2′脱氧胞苷处理人乳腺癌细胞株MDAMB435s,甲基化特异性PCR(MSP)检测Syk基因的甲基化,用逆转录聚合酶链反应(RTPCR)检测Syk基因的再表达,并用此处理过的细胞株和未经处理的细胞株分别接种于10裸鼠皮下,观察裸鼠成瘤率及肿瘤转移情况。结果MSP检测用5氮2′脱氧胞苷处理过的人乳腺癌MDAMB435s细胞株,Syk基因启动子没有甲基化,RTPCR可检测到Syk基因,用有Syk基因再表达的MDAMB435s细胞株接种于10只裸鼠皮下,8周后有2只形成肿瘤,无1例肺部转移,而无Syk基因表达细胞株10只都形成肿瘤,并在肺部形成转移灶。对照组的成瘤率及肺转移率显著高于治疗组。结论5氮2′脱氧胞苷通过去甲基化使MDAMB435细胞株中Syk基因再表达,Syk基因的再表达可抑制荷瘤裸鼠乳腺癌的生长和转移。
Objective To investigate the effect of re-expression of the spleen-derived tyrosine kinase (Syk) gene on the growth and metastasis of breast cancer in nude mice. Methods Methylation-specific PCR (MSP) was used to detect the methylation of Syk gene in A549 cells treated with nitrogen 2’-deoxycytidine. Syk gene expression was detected by reverse transcriptase-polymerase chain reaction (RTPCR) The treated cell lines and untreated cell lines were inoculated subcutaneously in 10 nude mice respectively to observe the tumor formation rate and tumor metastasis of nude mice. Results MSP detection of MDAMB435s cell line treated with 5-nitro-2’-deoxycytidine showed that Syk gene promoter was not methylated, Syk gene was detected by RTPCR, MDAMB435s cell line with Syk gene re-expression was inoculated into 10 Only the nude mice subcutaneously developed tumors after 2 weeks and none of the lung metastases. However, none of the 10 Syk gene-expressing cell lines formed tumors and formed metastases in the lung. The tumor formation rate and lung metastasis rate in the control group were significantly higher than those in the treatment group. Conclusion 5-deoxycytidine re-expression of Syk gene in MDAMB435 cell line by demethylation, Syk gene expression can inhibit the growth and metastasis of breast cancer in nude mice.