감마선 조사에 의한 뇌조직의 Jun 및 p53유전자 발현

Expression of Jun and p53 Genes from the Brain of Rats Irradiated with $^{60}Co{\gamma}$-ray

방사선은 DNA손상을 초래하고 세포 성장에 관련된 유전자의 발현 조절 및 apoptosis등을 유발한다고 알려져 있으며 본 연구는 신경계에 있어 방사선 조사 후 종양 발생율과 시간 경과의 관계 및 조사 양과 암 발생의 관계를 구명하기 위해 코발트 60의 전신조사에 따른 흰쥐 뇌 조직의 생체 반응을 연구하고자 하였다. 이를 위하여 상기조직의 jun 및 p53 유전자의 발현도를 1 Gy로 부터 100 Gy 범위의 감마선 용량별 및 1시간에서 6시간까지의 조사 훈 경과 시간 별로 Northern 분석하였다. Jun유전자 발현도는 ley이하에서 1시간 이내에 한계수준에 도달하였으며 30 Gy의 조사 1시간 째에 최대였다. 또한 조사 1시간 이후 1 Gy로부터 10 Gy 범위에서는 조사 5시간 및 6시간까지 점진적으로 증가되었으나 20 Gy로부터 100 Gy 범위에서는 조사 2시간까지 증가 후 감소되는 양상을 나타냈다. p53유전자의 발현도는 1 Gy이하에서 1시간 이내에 한계 수준에 근접했고 1 Gy의 조사 후 6시간 째에 최대였다. 1 Gy로부터 40 Gy까지의 범위에서는 조사 5시간 및 6시간까지 점진적으로 증가되는 반면 50 Gy에서 100 Gy범위에서는 조사 2시간 째까지 증가 후 감소되는 양상을 보였다. 따라서 감마선 조사양이 높을수록 jun 및 p53유전자는 신속하게 최대로 발현되었고 감마선 조사양이 낮을수록 서서히 증가되었다 그러나 jun유전자와 p53유전자의 감마선 조사에 따른 발현 양상에는 상호간의 연관성을 찾을 수 없었다.

Damage produced by radiation elicits a complex response in mammalian cells, including growth rate changes and the induction of a variety of genes associated with growth control and apoptosis. At doses of 10,000 cGy or greater, the exposed individual was killed in a matter of minutes to a couple of days, with symptoms consistent with pathology of the central nervous system(CNS) including degenerative changes. The nature of the damage in irradiated cells underlies the unique hazards of ionizing radiation. Radiation injury to CNS is a rare event in clinical medicine, but it is catastrophic for the patient in whom it occurs. The incidence of cerebral necrosis has been reported as high as 16% for doses greater than 6,000 cGy. In this study, the effect of radiation on brain tissue was studied in vivo. Jun and p53 genes in the rat brain were induced by whole body irradiation of rat with 600Co in doses between 1 Gy and 100 Gy and analyzed for expression of jun and p53 genes at the postirradiation time up to 6 hours. Northern analyses were done using 1.8 Kb & 0.8 Kb-pGEM-2-JUN/Eco RI/Pst I fragments, 2.0 Kb-php53B/Bam HI fragment and ,1.1 Kb-pBluescript SK--ACTIN/Eco RI fragment as the digoxigenin or [${\alpha}^{32}P$] dCTPlabeled probes for Jun, p53 and ${\beta}$-actin genes, respectively. Jun gene seemed to be expressed near the threshold levels in 1 hour after irradiation of $^{60}$Co in dose less than 1 Gy and was expressed in maximum at 1 hour after irradiation of $^{60}$Co in dose of 30 Gy. Jun was expressed increasingly with time until 5 or 6 hours after irradiation of $^{60}$Co in doses of 1 Gy and 10 Gy. After irradiation of $^{60}$Co in dose between 20 Gr and 100 Gy, the expression of Jun was however increased to peak in 2 hours and decreased thereafter. p53 gene in this study also seemed to be expressed near the threshold levels in 1 hour after irradiation of $^{60}$Co in dose less than 1 Gy and was expressed in maximum at 6 hours after irradiation of $^{60}$Co in dose of 1 Gy, p53 was expressed increasingly with time until 5 or 6 hours after irradiation of $^{60}$Co in dose between 1 Gy and 40 Gy. After irradiation of $^{60}$Co in doses of 50 Gy and 100 Gy, the expression of p53 was however increased to peak in 2 hours and decreased thereafter. The expression of Jun and p53 genes was not correlative in the brain tissue from rats. It seemed to be very important for the establishment of the optimum conditions for the animal studies relevant to the responses of genes inducible on DNA damage to ionizing radiation in mammalian cells. But there are many limitations to the animal studies such as the ununiform patterns of gene expression from the tissue because of its complex compositions. It is necessary to overcome the limitations for development of in situ Northern analysis.