Genome-wide DNA hypomethylation drives a more invasive pancreatic cancer phenotype and has predictive occult distant metastasis and prognosis potential
Genome-wide DNA hypomethylation drives a more invasive pancreatic cancer phenotype and has predictive occult distant metastasis and prognosis potential
复制标题
DOI：
10.3892/ijo.2022.5351
复制
发表时间：
2022-04
影响因子：
5.2
通讯作者：
Y. Endo;Koichi Suzuki;Yasuaki Kimura;Sawako Tamaki;Hidetoshi Aizawa;I. Abe;F. Watanabe;Takaharu Kato;Masaaki Saito;K. Futsuhara;H. Noda;F. Konishi;T. Rikiyama
中科院分区：
文献类型：
--
作者：
Y. Endo;Koichi Suzuki;Yasuaki Kimura;Sawako Tamaki;Hidetoshi Aizawa;I. Abe;F. Watanabe;Takaharu Kato;Masaaki Saito;K. Futsuhara;H. Noda;F. Konishi;T. Rikiyama
全基因组 DNA 低甲基化是与染色体不稳定 (CIN) 相关的人类癌症中最常见的分子特征，它参与调节胰腺癌 (PC) 转移的机制。体外研究了全基因组DNA低甲基化是否通过CIN影响PC的表型，并验证了其对PC生物学行为的意义。使用甲基化特异性定量 (q)PCR，利用人 PC 细胞系中长散布核苷酸元件 1 (LINE-1) 的相对去甲基化水平 (RDL) 来表征 DNA 低甲基化。使用比较基因组杂交分析，通过染色体拷贝数的变化来估计 CIN。通过免疫细胞化学评估染色体的异常分离，并使用抗γH2AX阳性细胞的数量评估DNA损伤反应。使用基质胶侵袭测定评估侵袭能力。对 49 名接受根治性手术的 PC 患者的临床样本进行了 DNA 低甲基化与临床结果的相关性评估。 PC 细胞中成功诱导全基因组 DNA 低甲基化导致特定染色体区域的拷贝数变化。随着抗γH2AX阳性细胞数量的增加，染色体异常分离的细胞数量显着增加。这些细胞的侵袭潜力也显着增加。临床标本中隐匿性远处转移的发生和接受者操作特征分析清楚地识别了那些可能和不可能发生隐匿性远处转移的患者，高LINE-1 RDL与隐匿性远处转移的存在（P=0.035）和不良预后（P=0.048）显着相关。揭示了全基因组 DNA 低甲基化对 PC 生物学行为的重要性，它在体外通过 CIN 促进更具侵袭性的表型，并预测 PC 患者对隐匿性远处转移的易感性和不良预后。
Genome-wide DNA hypomethylation is the most common molecular feature in human cancers associated with chromosomal instability (CIN), which is involved in the mechanisms that regulate pancreatic cancer (PC) metastasis. It was investigated whether genome-wide DNA hypomethylation affects the phenotype in PC via CIN in vitro, and its significance on the biological behavior of PC was verified. The relative demethylation level (RDL) of long interspersed nucleotide element-1 (LINE-1) in human PC cell lines was used to characterize DNA hypomethylation using methylation-specific quantitative (q)PCR. CIN was estimated by changes in chromosomal copy number using comparative genomic hybridization analysis. Abnormal segregation of chromosomes was assessed by immunocytochemistry, and the DNA damage response was evaluated using the number of anti-γH2AX positive cells. Invasion ability was assessed using a Matrigel invasion assay. Clinical specimens from 49 patients with PC who underwent curative surgery were evaluated for a correlation of DNA hypomethylation with clinical outcome. Successful induction of genome-wide DNA hypomethylation in PC cells led to copy number changes in specific chromosomal regions. The number of cells with abnormal segregation of chromosomes significantly increased with the number of anti-γH2AX positive cells. The invasive potential of these cells also significantly increased. The occurrence of occult distant metastasis in the clinical specimens and receiver operating characteristic analysis clearly identified those who were and were not likely to have occult distant metastasis, with high LINE-1 RDL significantly correlated with the presence of occult distant metastasis (P=0.035) and poor prognosis (P=0.048). The significance of genome-wide DNA hypomethylation on the biological behavior of PC, which promotes a more invasive phenotype via CIN in vitro and predicts the susceptibility to occult distant metastasis and poor prognosis in patients with PC was revealed.