Page 145 - D. Cancer biology

Suppression of MCF-7 Human Breast Cancer Cell Proliferation
by Globefish Homogenate

Junghoon Kim, and Jungho Kim

ABSTRACT

Breast cancer is one of the most aggressive cancers in women worldwide, including in South Korea. Although the treatment of this cancer has improved significantly, it is still the leading cause of
cancer‐related death in women. Previously, we reported that Globefish homogenate suppresses the growth of human colorectal cancer cells. To extend the applications of Globefish homogenate,
we investigated its cytotoxic effects on human breast cancer cells. Globefish homogenate was prepared by homogenizing 3-year-old Globefish. To assess the effects of Globefish homogenate on
growth of MCF-7 human breast cancer cells, cell proliferation and colony formation assays were performed using the cell counting and Crystal Violet staining methods. The 50% inhibitory
concentration (IC50) of Globefish homogenate on MCF-7 cell proliferation was calculated from the sigmoidal dose-response curve. Additionally, the cytotoxic role of Globefish homogenate was
confirmed by investigating major signaling pathways using Western blot analysis. The colony formation assay demonstrated that MCF-7 cells treated with Globefish homogenate formed up to
80% fewer colonies than control MCF-7 cells. Treatment with Globefish homogenate markedly suppressed the growth of MCF-7 cells in a dose-dependent manner. The sensitivity of the cells to
Globefish homogenate was determined by calculating the IC50; in this case, the IC50 was 210 ug/ml. Furthermore, significant downregulation of Cyclin D1 expression, along with phospho-Akt
and total Akt levels, was observed in MCF-7 cells treated with Globefish homogenate. This study demonstrates that treatment with Globefish homogenate inhibits the proliferation of MCF-7
human breast cancer cells by downregulating the expression of phosphor-Akt, total Akt, and Cyclin D1 proteins
RESULTS
IC50 of Globefish homogenate on MCF-7 human breast cancer cell proliferation
Inhibition of Colony forming effect in human breast cancer cells with Globefish Homogenate

Fig. 1. Inhibition of Colony forming effect in human breast cancer cells with Globefish Homogenate. (A) Crystal violet
staining of human breast cancer cell MCF at 3and 6day without or with 1% Homogenate. (B) Colony formation was
reduced with 1% Globe Homogenate, compared to Control at 6day. ** P < 0.01.

Effects of Globefish homogenate on growth of MCF-7 human breast cancer cells
Fig. 3. IC50 of Globefish homogenate on MCF-7 human breast cancer cell proliferation. (A)
A Representative images of MCF-7 human breast cancer cells with 0%, 0.001%, 0.01%, 0.1%, 1% Globe
Homogenate at 6days, Phase contrast, X100. (B) IC50 values of Globe Homogenate in MCF7 human
breast cancer cells. All values are expressed as means ± SD of 3 independent experiments, each done in
duplicates.
Downregulation of AKT in MCF7 human breast cancer cell with Globefish homogenate

B

Fig. 4. Downregulation of AKT in MCF7 human breast cancer cell with Globefish homogenate. (A) The
expression of phosphor-S473 Akt, Total Akt, CyclinD1 and loading control ß-actin at without or with 1% Globe
Homogenate, detected by Western blots. (B) Bar graph shows band density normalized to loading control and
compared to Control, VersaDoc MP 4000, All values are expressed as means ± SD of 3 independent
Fig. 2. Effects of Globefish homogenate on growth of MCF-7 human breast cancer cells (A) Representative experiments, * P <0.05, ** P < 0.01.
images of MCF7 Growth without or with 1% Globe Homogenate treatment for 6days, Phase contrast, X100.
Human breast cancer cell MCF7 was plated into 12-well plates at a density of 20000 cells/well. (B) The total
number of MCF-7 human breast cancer cells without or with 1% Globe homogenate was calculated every other
day for 6days. ** P < 0.01.
CONCLUSION
This study demonstrates that treatment with Globefish homogenate inhibits the proliferation of MCF-7 human breast cancer cells by downregulating the expression of phosphor-Akt, total Akt,
and Cyclin D1 proteins
REFERENCES

Kim J and Kim J. (2018). Inhibitory Effect of Globefish Homogenate on the Growth of Human Colorectal Cancer Cell Lines. KSBB Journal 33, 183-191
Contact information

Laboratory of Molecular and Cellular Biology, Department of Life Science, Sogang University, Seoul 40107, Korea
E-mail : jkim@sogang.ac.kr, Phone: 82-2-715-8461