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김재식,양선아,정경훈,김동윤,김광수,강현철 대한암학회 2023 Cancer Research and Treatment Vol.55 No.2
Purpose In the present study, we aimed to establish a liquid biopsy-based monitoring method using peripheral blood cell-free DNA (cfDNA) for patients with cervical cancer who underwent radical radiotherapy (RT). Materials and Methods Twenty-five patients with cervical cancer were prospectively recruited and treated with external beam RT and brachytherapy. In all patients, except one, chemotherapy was administered concurrently during RT. Whole peripheral blood samples were obtained at least twice from each patient. We performed next-generation sequencing (NGS) for the target-captured libraries (67 oncogenes and human papillomavirus [HPV] type 16/18) using 64 plasma cfDNA samples from the 25 participants. The ratio of HPV cfDNA and the variant allele frequency (VAF) in cfDNA was calculated, and their dynamic changes were monitored. The median follow-up duration was 25.4 months. Results In total, we identified 21,866 cfDNA variants. ARID1A and frameshift variants occupied the largest portion of altered genes and HIGH-grade variant types, respectively. In most cases, tumor shrinkage was followed by a decrease in the HPV ratio; however, an increase in HPV ratio indicated distant metastasis, despite the reduced tumor size. The initial HPV ratio reflecting the tumor burden was likely associated with treatment outcomes (p=0.16). We did not determine a role for serial changes in the VAF in cfDNA. Conclusion Our findings suggest that the HPV cfDNA ratio, calculated after targeted NGS, may be valuable for monitoring and predicting treatment responses. Accordingly, further validation of these findings is warranted.
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관세행정소송의 변화추이와 주요쟁점 : 대법원 판례 D/B를 중심으로
김재식 한국무역연구원 2013 무역연구 Vol.9 No.3
The purpose of this study is to find the trends and major issues of administrative litigation related to customs duties. For this study, the contents analysis method was adopted for the 93 precedents collected from the Supreme Court D/B. First, the number of lawsuits has been decreasing in the domains of collection, exemption and drawback. In the other hand, the number of legal conflicts has been maintaining the status quo in customs valuation and classification. Second, taxpayers often include the issue additionally which is the violation of the prohibition of retroactive taxation or the legal procedures for taxation by tax authorities. Third, some of the Supreme Court precedents play an important role in improving the related institutions. Fourth, the ratio of cases appealed by customs tax authorities is higher than the ratio by tax payers. In comparison, the winning ratio of customs tax authorities is a little bit lower. It should be noted that research results have the limitations of generalization and interpretation because this study is based only on the data base of the Supreme Court.
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김재식,이주호,전상록,김용섭,전승혁,우홍균 대한암학회 2023 Cancer Research and Treatment Vol.55 No.3
Purpose Exogenous epidermal growth factor (EGF) causes apoptosis in EGF receptor (EGFR)–overexpressing cell lines. The apoptosis-inducing factors could be a therapeutic target. We aimed to determine the mechanism of EGF-induced apoptosis using a genome-wide clustered regularly interspaced short palindromic repeats (CRISPR)-based knockout screen. Materials and Methods Two-vector system of the human genome-scale CRISPR knockout library v2 was used to target 19,050 genes using 123,411 single guide RNAs (sgRNAs). Recombinant human EGF (100 nM) or distilled water four times was administered to the experimental and control groups, respectively. The read counts of each sgRNA obtained from next-generation sequencing were analyzed using the edgeR algorithm. We used another EGFR-overexpressing cell line (A549) and short hairpin RNAs (shRNAs) targeting five EGF-resistance genes for validation. DUSP1 expression in A431, A549, and HEK293FT cells was calculated using reverse transcription–quantitative polymerase chain reaction. Results We found 77 enriched and 189 depleted genes in the experimental group using the CRISPR-based knockout screen and identified the top five EGF-resistance genes: DDX20, LHFP, REPS1, DUSP1, and KRTAP10-12. Transfecting shRNAs targeting these genes into A549 cells significantly increased the surviving fractions after EGF treatment, compared with those observed in the control shRNA-transfected cells. The expression ratio of DUSP1 (inhibits ERK signaling) increased in A431 and A549 cells after EGF treatment. However, DUSP1 expression remained unchanged in HEK293FT cells after EGF treatment. Conclusion The CRISPR-based knockout screen revealed 266 genes possibly responsible for EGF-induced apoptosis. DUSP1 might be a critical component of EGF-induced apoptosis and a novel target for EGFR-overexpressing cancers.
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