关键词: Cell morphology DNA fragmentation immunocytochemical staining liquid-based cytology pancreatic cancer real-time PCR

来  源:   DOI:10.4103/joc.joc_13_22   PDF(Pubmed)

Abstract:
UNASSIGNED: Liquid-based cytology (LBC) has been widely used since 2000. Next-Generation Sequencing (NGS) analysis of residual specimens in LBC fixative may also be performed for pancreatic cancer in the near future. We examined cell morphology, antigenicity and nucleic acids in pancreatic cancer cells at different fixation times using two types of LBC fixatives.
UNASSIGNED: PANC-1 cells were fixed in 1 ml CytoRich Red (CR), CytoRich Blue (CB), 95% ethanol (95% AL) or 10% neutral buffered formalin (10% NBF) and evaluated for cell area, antigenicity and nucleic acids with fixation times of 1 hour and 1, 3, 9, and 14 days. Antigenicity was evaluated by immunocytochemical staining for p53 and CK20, and nucleic acid fragmentation was assessed by real-time PCR.
UNASSIGNED: There was no difference in total cell area between 1 hour and 14 day fixation times for the CR group, but the CB group showed cell contraction with 9 days fixation. In immunocytochemical staining, the CR group showed high p53 and CK20 positivity even after 14 days fixation. The CB group had a lower p53 positive rate than the CR group from 1 hour fixation. For nucleic acid fragmentation, Ct values for the CR group increased with fixation time. The CB group had consistently low Ct values.
UNASSIGNED: Different LBC fixatives and fixation time can have varying effects on cell morphology, antigenicity and nucleic acids in pancreatic cancer cells. Therefore, fixative type and fixation time should be considered for molecular testing on residual samples in LBC fixatives.
摘要:
自2000年以来,液基细胞学(LBC)已被广泛使用。LBC固定剂中残留标本的下一代测序(NGS)分析也可能在不久的将来用于胰腺癌。我们检查了细胞形态,使用两种类型的LBC固定剂在不同固定时间的胰腺癌细胞中的抗原性和核酸。
将PANC-1细胞固定在1ml富细胞红(CR)中,富细胞蓝(CB),95%乙醇(95%AL)或10%中性缓冲福尔马林(10%NBF),并评估细胞面积,抗原性和核酸,固定时间为1小时和1、3、9和14天。通过p53和CK20的免疫细胞化学染色评估抗原性,并通过实时PCR评估核酸片段化。
在1小时和14天固定时间之间,CR组的总细胞面积没有差异,但CB组显示细胞收缩,固定9天。在免疫细胞化学染色中,即使在固定14天后,CR组仍显示出高p53和CK20阳性。从1小时固定开始,CB组的p53阳性率低于CR组。对于核酸片段化,CR组的Ct值随固定时间增加。CB组的Ct值始终较低。
不同的LBC固定剂和固定时间会对细胞形态产生不同的影响,胰腺癌细胞中的抗原性和核酸。因此,在对LBC固定剂中残留样品进行分子检测时,应考虑固定剂类型和固定时间.
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