Abstract：
BACKGROUND: Altered glycosylation plays a role in carcinogenesis. GALNT14 promotes cancer stem-like properties and drug resistance. GDF-15 is known to induces drug resistance and stemness markers for maintenance of breast cancer (BC) stem-like cell state. Currently there is lack of data on association of GDF-15 and GALNTs. In this study, the expression and interaction of GALNT14 and GDF-15 with stemness (OCT4 and SOX2) and drug resistance (ABCC5) markers were evaluated in BC.
METHODS: We investigated tumour tissue from 30 BC patients and adjacent non-tumour tissues. Expression of serum GALNT14 from BC patients and matched healthy controls was evaluated. Expression of GALNT14, GDF-15, OCT4, SOX2, ABCC5, and β-catenin in BC tissue was determined by RT-PCR. Knockdown of GALNT14 and GDF-15 in the MCF-7 cell line was done through siRNA, gene expression and protein expression of β-catenin by western blot were determined.
RESULTS: A significant increase in the expression of GALNT14, GDF-15, OCT4, SOX2, ABCC5, and β-catenin was observed in BC tumour tissues compared to adjacent non-tumour tissues. The serum level of GALNT14 was significantly high in BC patients (80.7 ± 65.3 pg/ml) compared to healthy controls (12.2 ± 9.12 pg/ml) (p < 0.000). To further analyse the signalling pathway involved in BC stemness and drug resistance, GALNT14 and GDF-15 were knocked down in the MCF-7 cell line, and it was observed that after knockdown, the expression level of OCT4, SOX2, ABCC5, and β-catenin was decreased, and co-knockdown with GALNT14 and GDF-15 further decreased the expression of genes.
CONCLUSIONS: It can be concluded that GALNT14, in association with GDF-15, promotes stemness and intrinsic drug resistance in BC, possibly through the β-catenin signalling pathway.
METHODS: We investigated tumour tissue from 30 BC patients and adjacent non-tumour tissues. Expression of serum GALNT14 from BC patients and matched healthy controls was evaluated. Expression of GALNT14, GDF-15, OCT4, SOX2, ABCC5, and β-catenin in BC tissue was determined by RT-PCR. Knockdown of GALNT14 and GDF-15 in the MCF-7 cell line was done through siRNA, gene expression and protein expression of β-catenin by western blot were determined.
RESULTS: A significant increase in the expression of GALNT14, GDF-15, OCT4, SOX2, ABCC5, and β-catenin was observed in BC tumour tissues compared to adjacent non-tumour tissues. The serum level of GALNT14 was significantly high in BC patients (80.7 ± 65.3 pg/ml) compared to healthy controls (12.2 ± 9.12 pg/ml) (p < 0.000). To further analyse the signalling pathway involved in BC stemness and drug resistance, GALNT14 and GDF-15 were knocked down in the MCF-7 cell line, and it was observed that after knockdown, the expression level of OCT4, SOX2, ABCC5, and β-catenin was decreased, and co-knockdown with GALNT14 and GDF-15 further decreased the expression of genes.
CONCLUSIONS: It can be concluded that GALNT14, in association with GDF-15, promotes stemness and intrinsic drug resistance in BC, possibly through the β-catenin signalling pathway.
摘要：
背景：糖基化改变在致癌作用中起作用。GALNT14促进癌症干细胞样特性和耐药性。已知GDF-15诱导用于维持乳腺癌(BC)干细胞样细胞状态的耐药性和干性标志物。目前缺乏关于GDF-15和GALNT的关联的数据。在这项研究中,在BC中评估了GALNT14和GDF-15与干性（OCT4和SOX2）和耐药性（ABCC5）标记的表达和相互作用。
方法：我们研究了30例BC患者的肿瘤组织和邻近的非肿瘤组织。评估来自BC患者和匹配的健康对照的血清GALNT14的表达。通过RT-PCR测定BC组织中GALNT14，GDF-15，OCT4，SOX2，ABCC5和β-catenin的表达。通过siRNA对MCF-7细胞系中的GALNT14和GDF-15进行敲除，蛋白质印迹法测定β-catenin的基因表达和蛋白表达。
结果：与邻近的非肿瘤组织相比，在BC肿瘤组织中观察到GALNT14，GDF-15，OCT4，SOX2，ABCC5和β-catenin的表达显着增加。与健康对照组（12.2±9.12pg/ml）相比，BC患者的GALNT14血清水平显着升高（80.7±65.3pg/ml）（p<0.000）。为了进一步分析BC干性和耐药性的信号通路，GALNT14和GDF-15在MCF-7细胞系中被敲低，据观察，在击倒后，OCT4，SOX2，ABCC5和β-catenin的表达水平降低，与GALNT14和GDF-15的共敲除进一步降低了基因的表达。
结论：可以得出结论，GALNT14与GDF-15相关，可促进BC的干性和内在耐药性，可能通过β-连环蛋白信号通路。
方法：我们研究了30例BC患者的肿瘤组织和邻近的非肿瘤组织。评估来自BC患者和匹配的健康对照的血清GALNT14的表达。通过RT-PCR测定BC组织中GALNT14，GDF-15，OCT4，SOX2，ABCC5和β-catenin的表达。通过siRNA对MCF-7细胞系中的GALNT14和GDF-15进行敲除，蛋白质印迹法测定β-catenin的基因表达和蛋白表达。
结果：与邻近的非肿瘤组织相比，在BC肿瘤组织中观察到GALNT14，GDF-15，OCT4，SOX2，ABCC5和β-catenin的表达显着增加。与健康对照组（12.2±9.12pg/ml）相比，BC患者的GALNT14血清水平显着升高（80.7±65.3pg/ml）（p<0.000）。为了进一步分析BC干性和耐药性的信号通路，GALNT14和GDF-15在MCF-7细胞系中被敲低，据观察，在击倒后，OCT4，SOX2，ABCC5和β-catenin的表达水平降低，与GALNT14和GDF-15的共敲除进一步降低了基因的表达。
结论：可以得出结论，GALNT14与GDF-15相关，可促进BC的干性和内在耐药性，可能通过β-连环蛋白信号通路。
