Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Cytotoxicity of Particulate Matter in Various Human Cells Lines

미세먼지가 다양한 사람 세포주에 미치는 세포 독성

  • Lee, Ji-Hyeon (Science Education Center for the Gifted Gyeongsang National University) ;
  • Lee, Joo-Yeong (Science Education Center for the Gifted Gyeongsang National University) ;
  • Kim, Mi-Jeong (Department of Biology Education, Gyeongsang National University) ;
  • Kim, Hyeon-Ji (Department of Biology Education, Gyeongsang National University) ;
  • Lee, Sung-Ho (Division of Life Science, Gyeongsang National University) ;
  • Jeon, Byeong-Gyun (Science Education Center for the Gifted Gyeongsang National University)
  • 이지현 (경상대학교 과학영재교육원) ;
  • 이주영 (경상대학교 과학영재교육원) ;
  • 김미정 (경상대학교 사범대학 생물교육과) ;
  • ;
  • 이성호 (경상대학교 생명과학부) ;
  • 전병균 (경상대학교 과학영재교육원)
  • Received : 2019.02.13
  • Accepted : 2019.05.29
  • Published : 2019.06.30
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Abstract

The present study investigated the cytotoxicity of particulate matter (PM) derived from car air filter (outdoor PM) and home cleaner filter (indoor PM) in the various human cell lines. Each outdoor and indoor PM were harvested by ethanol extraction method, subsequently sieved with 10 um filter paper, sterilized with autoclave and added to culture media. The half maximal inhibitory concentration ($IC_{50}$) values was significantly (p<0.05) lower in the outdoor PM, compared with indoor PM, and the significantly (p<0.05) higher $IC_{50}$ values were observed in the cancer cell lines (A-549 lung adenocarcinoma and AGS stomach adenocarcinoma), than those of normal MRC-5 fibroblasts and dental papilla tissue derived-mesenchymal stem cells (DSC). After being exposed to $100{\mu}g/ml$ outdoor PM for 7 days, the population doubling time (PDT) was significantly (p<0.05) increased in especially MRC-5 and DSC cell lines, compared with untreated cell lines. Further, the expression of senescence-associated ${\beta}$-galactosidase activity was up-regulated in all the cells exposed to outdoor PM than those of untreated control. Besides, the expression level of inflammation-associated genes, such as cyclooxygenase-2 (COX-2) and interleukin-6 (IL-6) was found to be significantly (p<0.05) increased in the outdoor PM-treated cell lines than those of untreated cell lines. Our results showed that PM induces the cytotoxicity via arrest of cell growth, cell damage and inflammation response.

본 연구는 대기 중에 떠다니는 입자인 미세먼지의 발생원에 따라 여러 종류의 사람 세포주에 미치는 세포 독성 효과를 검증해 보고자 하였다. 실험에 사용된 미세먼지는 자동차의 공기 필터(차 미세먼지, 실외)와 집에 있는 청소기의 필터(집 미세먼지, 실내)에서 유래한 미세먼지를 에탄올 추출법으로 포집하여 여과한 다음 대략 $10{\mu}m$ 이하의 미세먼지를 세포배양액에 첨가하였다. MTT 분석 방법으로 조사된 세포성장의 반억제농도 값($IC_{50}$)은 집 미세먼지보다 차 미세먼지에서 각 세포에 대한 $IC_{50}$ 값이 유의적으로(p<0.05) 더 낮았고, 정상세포주인 섬유아세포(MRC-5) 및 사랑니 유래 중간엽성 성체줄기세포(DSC)에서 $IC_{50}$ 값은 폐암세포주(A-549) 및 위암세포주(AGS)에 비해 유의적으로(p<0.05) 더 낮았다. 차 미세먼지를 $100{\mu}g/ml$의 농도로 첨가하여 1주일동안 세포를 배양하여 세포배가시간을 조사하였던 바, 암세포주보다 MRC-5 및 DSC 세포주에서 미세먼지의 처리 후 세포배가시간이 유의적으로(p<0.05) 늘어나는 것을 관찰하였고, 미세먼지에 노출된 세포는 노화 관련 베타-갈락토시다아제의 발현이 증가하여 세포의 노화가 일어나는 것을 관찰하였다. 또한, 차 미세먼지를 각 세포주의 $IC_{50}$ 값으로 1주일 동안 처리한 후, 염증 관련 유전자인 COX-2 및 IL-6의 발현이 유의적으로(p<0.05) 증가하는 것을 관찰하였다. 이상의 결과로 보아, 미세먼지는 세포의 성장을 억제하고, 손상을 일으키면서 염증의 발현을 유도하는 것으로 조사되었다.

Keywords

SMGHBM_2019_v29n6_724_f0001.png 이미지

Fig. 1. Collection of particulate matter derived from car air filter.

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Fig. 2. Cell morphology in DSC and AGS cell lines treated with particulate matter under inverted microscope.

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Fig. 3. Analysis of cytotoxicity and determination of IC50 values by MTT assay in MRC-5, DSC, A-549 and AGS exposed to particulate matter.

SMGHBM_2019_v29n6_724_f0004.png 이미지

Fig. 4. Analysis of population doubling time in each cell line treated with 100 μg/ml outdoor particulate matter.

SMGHBM_2019_v29n6_724_f0005.png 이미지

Fig. 5. Analysis of senescence-associated-β-galactosidase activity in MRC-5 and DSC treated with 100 μg/ml outdoor particulate matter.

SMGHBM_2019_v29n6_724_f0006.png 이미지

Fig. 6. Analysis of senescence-associated-β-galactosidase activity in A-549 and AGS treated with 100 μg/ml and 1,000 μg/ml outdoor particulate matter, respectively.

SMGHBM_2019_v29n6_724_f0007.png 이미지

Fig. 7. Expression analysis of COX-2 and IL-6 transcripts related with inflammation in each cell line treated with particulate matter.

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