• Biomolecules & Therapeutics
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• 제22권2호
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• pp.114-121
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• 2014

Refractoriness of acute myeloid leukemia (AML) cells to chemotherapeutics represents a major clinical barrier. Suicide gene therapy for cancer has been attractive but with limited clinical efficacy. In this study, we investigated the potential application of herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) based system to inhibit chemoresistant AML cells. We first generated Ara-C resistant K562 cells and doxorubicin-resistant THP-1 cells. We found that the HSV-TK/GCV anticancer system suppressed drug resistant leukemic cells in culture. Chemoresistant AML cell lines displayed similar sensitivity to HSV-TK/GCV. Moreover, HSV-TK/GCV killing of leukemic cells was augmented to a mild but significant extent by all-trans retinoic acid (ATRA) with concomitant upregulation of Connexin 43, a major component of gap junctions. Interestingly, HSV-TK/GCV killing was enhanced by expression of vesicular stomatitis virus G glycoprotein (VSV-G), a fusogenic membrane protein, which also increased leukemic cell fusion. Co-culture resistant cells expressing HSV-TK and cells stably transduced with VSV-G showed that expression of VSV-G could promote the bystander killing effect of HSV-TK/GCV. Furthermore, combination of HSV-TK/GCV with VSV-G plus ATRA produced more pronounced antileukemia effect. These results suggest that the HSV-TK/GCV system in combination with fusogenic membrane proteins and/or ATRA could provide a strategy to mitigate the chemoresistance of AML.

• 생약학회지
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• 제32권4호통권127호
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• pp.274-279
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• 2001

Ailanthus altissima belonging to the family Simaroubaceae has been used to settle an upset stomach, to combat a fever, and as an insecticide. Apoptosis is an active process, which is a critical feature of the regulated development of multicellular organisms. We investigated whether the extract of A. altissima induced apoptotic cell death in Jurkat T-acute lymphoblastic leukemia (ALL) cells. Upon treatments with the extract, the dose-dependent inhibitions of cell viability were observed. It also caused apoptosis as measured by cell morphology and DNA fragmentation. The capability of the extract to induce apoptosis was associated with proteolytic cleavage of specific target protein such as poly(ADP-ribose)polymerase (PARP) protein, suggesting the possible involvement of the activations of caspases. Further study showed that Bcl-2 protein levels were not changed in all treated groups compared to control group. These results suggest that A. altissima induces Bcl- 2-independent apoptosis in Jurkat T cells.

• Advances in Traditional Medicine
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• 제5권1호
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• pp.21-28
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• 2005

The leaf extracts of Araucaria bidwillii Hook. (Araucariaceae) were evaluated for their cytotoxic effect in various human cancer cell lines. Preliminary investigation by brine shrimp lethality assay indicated that $LC_{50}$ value of various successive extracts were found to be less than $1000\;{\mu}g/ml$, where the ethyl acetate extract showed maximum activity of less than $100\;{\mu}g/ml$. Further cytotoxic evaluation of various leaf extracts of Araucaria bidwilli Hook was carried out in four different human cancer cell lines-acute myeloblastic leukemia (HL-60), chronic myelogenic leukemia (K-562), breast adenocarcinoma (MCF-7) and cervical epithelial carcinoma (HeLa). Cytotoxicity was assessed by trypan blue dye exclusion method and 3-(4,5-dimethyl thiazole-2yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay. From the present investigation it was found that the ethyl acetate and methanol extract of Araucaria bidwilli Hook was found to be more effective in leukemic cell lines and was less effective in MCF-7 and HeLa. The $IC_{50}$ value of the ethyl acetate extract in leukemic cell lines was found to be $28.18\;and\;34.64\;{\mu}g/ml$ and methanol extract was found to be $33.11\;&\;39.81\;{\mu}g/ml$. It can be concluded that various extract from the leaves of Araucaria bidwillii Hook. posses cytotoxic activity tested in brine shrimps and various human carcinoma cell lines.

• Asian Pacific Journal of Cancer Prevention
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• 제16권15호
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• pp.6663-6668
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• 2015

Background: Treatment failure in leukemia is due to either pharmacokinetic resistance or cell resistance to drugs. Materials and Methods: Gene expression of multiple drug resistance protein (MDR-1), multidrug resistance-related protein (MRP) and low resistance protein (LRP) was assessed in 45 pediatric ALL cases and 7 healthy controls by real time PCR. The expression was scored as negative, weak, moderate and strong. Results: The male female ratio of cases was 2.75:1 and the mean age was 5.2 years. Some 26/45 (58%) were in standard risk, 17/45(38%) intermediate and 2/45 (4%) in high risk categorie, 42/45 (93%) being B-ALL and recurrent translocations being noted in 5/45 (11.0%). Rapid early response (RER) at day 14 was seen in 37/45 (82.3%) and slow early response (SER) in 8/45 (17.7%) cases. Positive expression of MDR-1, LRP and MRP was noted in 14/45 (31%), 15/45 (33%) and 27/45 (60%) cases and strong expression in 3/14 (21%), 11/27 (40.7%) and 8/15 (53.3%) cases respectively. Dual or more gene positivity was noted in 17/45 (38%) cases. 46.5 % (7/15) of LRP positive cases at day 14 were in RER as compared to 100% (30/30) of LRP negative cases (p<0.05). All 8 (100%) LRP positive cases in SER had strong LRP expression (p=<0.05). Moreover, only 53.3% of LRP positive cases were in haematological remission at day 30 as compared to 100% of LRP negative cases (p=<0.05). Conclusions: Our study indicated that increased LRP expression at diagnosis in pediatric ALL predicts poor response to early treatment and hence can be used as a prognostic marker. However, larger prospective studies with longer follow up are needed, to understand the clinical relevance of drug resistance proteins.

• 동의생리병리학회지
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• 제19권4호
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• pp.903-907
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• 2005

Many naturally occurring plant extracts are studied for their beneficial effects for health and particularly on cancer. Apoptosis, or programmed cell death, occurs in both normal and pathological conditions, including cancer Dysregulation of apoptosis allows transformed cells to continually and uninhibitedly enter the cell cycle, thus perpetuating the sequence of mutation, genomic instability and, finally, oncogenesis. To investigate the apoptosis-inducing effect of the extract of Fructus Trichosanthis (EFT) on leukemic HL-60 cells and its mechanism, HL-60 cells in vitro in culture medium were given different doses of the extract. The inhibitory rate of cells were measured by microculture tetrazolium assay, cell apoptotic rate was detected by flow cytometry, morphology of cell apoptosis was observed by DAPI fluorescence staining, and the activations of caspases and PARP were detected using Western blotting analysis. The extract could activate the caspase-3 and caspase-8, induce PARP cleavage, inhibit growth of HL-60 cells, and cause apoptosis significantly The suppression was in dose-dependent manner. Marked morphological changes of cell apoptosis including condensation of chromatin and nuclear fragmentation were observed clearly by DAPI fluorescence staining especially. These results will provide strong laboratory evidence of EFT for clinical treatment of acute leukemia.

• Journal of Korean Neurosurgical Society
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• 제60권5호
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• pp.534-539
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• 2017

Objective : Patients with hematological malignancies frequently encounter spine-related symptoms, which are caused by disease itself or process of treatment. However, there is still lack of knowledge on their epidemiology and clinical courses. The purpose of this article is to review clinical presentations and surgical results for spinal involvement of hematologic malignancies. Methods : From January 2011 to September 2014, 195 patients (98 males and 97 females) suffering from hematological malignancies combined with spinal problems were retrospectively analyzed for clinical and radiological characteristics and their clinical results. Results : The most common diagnosis of hematological malignancy was multiple myeloma (96 patients, 49.7%), followed by chronic myeloid leukemia (30, 15.2%), acute myeloid leukemia (22, 11.2%), and lymphoma (15, 7.56%). The major presenting symptoms were mechanical axial pain (132, 67.7%) resulting from pathologic fractures, and followed by radiating pain (49, 25.1%). Progressive neurologic deficits were noted in 15 patients (7.7%), which revealed as cord compression by epidural mass or compressive myelopathy combined with pathologic fractures. Reconstructive surgery for neurologic compromise was done in 16 patients. Even though surgical intervention was useful for early paralysis (Frankel grade D or E), neurologic recovery was not satisfactory for the progressed paralysis (Frankel grade A or B). Conclusion : Hematological malignancies may cause various spinal problems related to disease progression or consequences of treatments. Conservative and palliative treatments are mainstay for these lesions. However, timely surgical interventions should be considered for the cases of pathologic fractures with progressive neurologic compromise.

• 한국전통의학지
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• 제15권1호
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• pp.83-89
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• 2006

Many naturally occurring plant extracts are studied for their beneficial effects for health and particularly on cancer. Apoptosis, or programmed cell death, occurs in both normal and pathological conditions, including cancer. Dysregulation of apoptosis allows transformed cells to continually and uninhibitedly enter the cell cycle, thus perpetuating the sequence of mutation, genomic instability and, finally, oncogenesis. To investigate the apoptosis-Inducing effect of the extract of Fructus Trichosanthis (EFT) on leukemic HL-60 cells and its mechanism, HL-60 cells in vitro in culture medium were given different doses of the extract. The inhibitory rate of cells were measured by microculture tetrazolium assay, cell apoptotic rate was detected by flow cytometry, morphology of cell apoptosis was observed by DAPI fluorescence staining, and the activations of caspases and PARP were detected using Western blotting analysis. The extract could activate the caspase-3 and caspase-8, induce PARP cleavage, inhibit growth of HL-60 cells, and cause apoptosis significantly. The suppression was in dose-dependent manner. Marked morphological changes of cell apoptosis including condensation of chromatin and nuclear fragmentation were observed clearly by DAPI fluorescence staining especially. These results will provide strong laboratory evidence of EFT for clinical treatment of acute leukemia.

• Clinical and Experimental Pediatrics
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• 제51권1호
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• pp.73-77
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• 2008

목 적 : p16 유전자는 염색체 9p21에 위치하는데 종양억제 유전자 중 하나로 cyclin-dependent kinase의 억제제로 작용하며 Rb 인산화를 억제한다. 다양한 종류의 종양에서 p16 유전자의 결실 또는 과메틸화가 발견되고 있는데 이는 급성림프구성백혈병에서도 흔히 발견되는 이상 소견으로 높은 빈도로 나타나고 있지만 급성림프구성백혈병의 예후와 p16 유전자의 연관성에 대해서는 아직 논란의 여지가 있다. 본 연구에서는 면역조직화학염색으로 확인한 p16 단백질의 소실과 급성림프구성백혈병 환아들의 임상 경과와의 연관성에 대하여 조사하고자 하였다. 방 법 : 1998년 1월부터 2006년 12월까지 급성림프구성백혈병으로 진단된 74명의 진단 시 골수 슬라이드에서 p16 단백질 면역조직화학염색을 하였다. 환아들의 임상 양상, 검사실 소견, 치료 후 경과에 대해서 후향적으로 조사하였다. 결 과 : 74명 중 12명에서 p16 단백질이 면역화학염색 결과 음성이었다. 이들 중 남아가 7명 여아가 5명이었으며 진단 시 연령의 중앙값은 5.8(1.3-18.8)세였다. 백혈구 수의 중앙값은 17,225 $(500-403,300)/{\mu}L$ 이었으며 면역표현형은 early pre-B CALLA (+)형이 7명, T 세포형은 5명이었다. 진단 시 예후 중간군이었던 두 명의 환아들에서 골수 재발 하였으며 3명의 환아들이 유방암의 가족력이 있었다. 4명이 사망하여 8년 생존율은 $53.5{\pm}18.7%$였다. 결 론 : p16 단백질의 소실은 소아 급성림프구성백혈병에서 불량한 예후와 연관된 인자로 추정되며 임상에서 진단 시 p16에 대한 유전자 검사뿐만 아니라 단백질에 대해서도 검사가 필요할 것으로 생각된다. 하지만 좀더 많은 환자들에 대한 분석이 더 정확한 연관성을 밝히는데 도움이 될 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5469-5475
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• 2012

Background and Objectives: Acute lymphoblastic leukemia (ALL) is a complex genetic disease involving many fusion oncogenes (FO) having prognostic significance. The frequency of various FO can vary in different ethnic groups, with important implications for prognosis, drug selection and treatment outcome. Method: We studied fusion oncogenes in 101 pediatric ALL patients using interphase FISH and RT-PCR, and their associations with clinical features and treatment outcome. Results: Five most common fusion genes i.e. BCR-ABL t (22; 9), TCF3-PBX1 (t 1; 19), ETV6-RUNX1 (t 12; 21), MLL-AF4 (t 4; 11) and SIL-TAL1 (del 1p32) were found in 89/101 (88.1%) patients. Frequency of BCR-ABL was 44.5% (45/101). BCR-ABL positive patients had a significantly lower survival ($43.7{\pm}4.24$ weeks) and higher white cell count as compared to others, except patients with MLL-AF4. The highest relapse-free survival was documented with ETV6-RUNX1 (14.2 months) followed closely by those cases in which no gene was detected (13.100). RFS with BCR-ABL, MLL-AF4, TCF3-PBX1 and SIL-TAL1 was less than 10 months (8.0, 3.6, 5.5 and 8.1 months, respectively). Conclusions: This is the first study from Pakistan correlating molecular markers with disease biology and treatment outcome in pediatric ALL. It revealed the highest reported frequency of BCR-ABL FO in pediatric ALL, associated with poor overall survival. Our data indicate an immediate need for incorporation of tyrosine kinase inhibitors in the treatment of BCR-ABL+ pediatric ALL in this population and the development of facilities for stem cell transplantation.

• Clinical and Experimental Pediatrics
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• 제50권12호
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• pp.1217-1224
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• 2007

목 적 : 소아 ALL 관해유도 시 스테로이드는 중추적인 역할을 하는 약제로 덱사메타손은 프레드니솔론에 비해 항백혈병 효과가 우수하지만 감염, 혈당 불내성, 골 괴사 등의 합병증의 가능성도 높다. 본원에서는 백혈병 세포에 대한 높은 항암효과를 고려하여 관해유도 시 덱사메타손을 사용했지만 이 약제에 의한 합병증 감소를 위해 2005년부터 시행한 치료지침(CMCPL-2005)에는 덱사메타손의 투여기간을 총 4주에서 3주로 일주일 단축해서 투여하였다. 이에 저자들은 수정한 치료지침의 관해유도율 및 주요 합병증 발생률을 기존지침(CMCPL-2001)의 성적과 비교 분석하기로 하였다. 방 법 : 가톨릭대학교 성모병원 소아과에서 2001년 1월부터 2006년 12월까지 ALL로 진단 받은 환아들의 기록을 후향적으로 조사하였다. 2001년 1월부터 2004년 12월까지 기간 동안 ALL을 진단받은 환아들은 CMCPL-2001지침 하에 관해유도를 받았고 2005년 이후에 진단받은 환아들은 CMCPL-2005지침 하에 관해유도를 받았다. 대상 환아에 대한 기초자료로 나이, 성별, 진단 시 백혈구 수, 면역표현형, 세포유전학적 특징, 그리고 위험군 분류에 대해 조사하였다. 각 CMCPL-2001 및 CMCPL-2005군의 전처치 기간 동안의 스테로이드 투여에 대한 반응을 확인하고 관해유도 결과를 분석하였다. 관해유도를 위해 입원한 기간 동안 발생한 감염, 간독성, 신경계, 위장관계, 내분비계 합병증 등은 NCI 독성 기준분류를 토대로 조사하였다. 결 과 : 총 141명이 CMCPL-2001지침으로, 88명이 CMCPL- 2005지침으로 관해유도를 받았다. 고위험군에 해당하는 10세 이상 환아, 예후 불량인자에 포함되는 염색체전위가 CMCPL-2005군에 유의하게 많았다. CMCPL-2001군에서는 총 132례(94%), CMCPL-2005군에서는 81례(92%)가 전처치 스테로이드에 양호한 반응을 보여 통계적인 차이는 없었고 관해유도 치료 후 시행한 골수검사에서도 CMCPL-2001군에서는 136례(96%), CMCPL- 2005군에서는 82례(93%)가 완전관해를 보여 동등한 치료성적을 보였다. CMCPL-2005군이 신경계, 위장관계, 심혈관계 등의 스테로이드와 연관된 합병증 없이 관해유도를 받을 확률이 CMCPL- 2001군보다 높았지만 감염성 합병증과 같은 중증합병증에 대해서는 두 군간에 차이가 없었다. 결 론 : 소아 ALL의 관해유도 치료지침에서 덱사메타손의 투여를 일주일 단축해도 기존의 4주간 스테로이드 투여방법과 동등한 관해 유도율을 확인하였으나 감염성 합병증 등 스테로이드 투여와 연관된 합병증의 감소는 없었다.