Calinisan Joan H, Chan Steven R, King Alan, Chan Philip J
Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, California 92350, USA.
J Assist Reprod Genet. 2002 Mar;19(3):132-6. doi: 10.1023/a:1014736805127.
The effect of human papillomavirus (HPV) DNA from the E6-E7 region on the integrity of DNA in blastocyst stage embryonic cells was studied. The study design paralleled the event whereby HPV DNA from the infecting virus would target host cell DNA. The objectives were (a) to determine if the DNA of blastocysts were disrupted by the presence of HPV DNA and (b) to determine if the intensity of DNA damage was associated with the type of HPV.
This study involved superovulating female mice, mating, collecting one-cell embryos, and culturing to the expanded blastocyst stage. The blastocysts were infected with PCR-synthesized DNA fragments from either HPV type 16, 18, 31, or 33. The blastocyst DNA were analyzed by comet assay after 24 h of incubation. The fluorescent images were digitized and the pixel intensity of each blastocyst was measured.
Only the DNA of HPV type 16 was associated with significant DNA fragmentation in comparison with the other HPV types. There was no relationship between HPV DNA fragment size and the intensity of DNA fragmentation.
The data suggested that one mode of action of HPV type 16 was to initiate apoptosis of embryonic cells through DNA fragmentation. The effect of HPV 16 occurred rapidly within 24 h. The intensity of DNA damage was not linked to the specific type of HPV. However, the results do not rule out the other HPV types affecting embryos under conditions different from this study.
研究来自E6 - E7区域的人乳头瘤病毒(HPV)DNA对囊胚期胚胎细胞中DNA完整性的影响。该研究设计模拟了感染病毒中的HPV DNA靶向宿主细胞DNA的过程。目标是:(a)确定HPV DNA的存在是否会破坏囊胚的DNA;(b)确定DNA损伤的强度是否与HPV类型有关。
本研究涉及对雌性小鼠进行超排卵、交配、收集单细胞胚胎并培养至扩张囊胚期。将囊胚用PCR合成的来自16型、18型、31型或33型HPV的DNA片段进行感染。孵育24小时后,通过彗星试验分析囊胚DNA。将荧光图像数字化并测量每个囊胚的像素强度。
与其他HPV类型相比,只有16型HPV的DNA与显著的DNA片段化有关。HPV DNA片段大小与DNA片段化强度之间没有关系。
数据表明,16型HPV的一种作用方式是通过DNA片段化引发胚胎细胞凋亡。16型HPV的作用在24小时内迅速发生。DNA损伤的强度与HPV的特定类型无关。然而,结果并不排除在与本研究不同的条件下其他HPV类型对胚胎产生影响。