In Vivo-Like Cell-Cycle Phase Distribution of Cancer Cells in Gelfoam® Histoculture Observed in Real Time by FUCCI Imaging

The objective was to determine the antiproliferative and apoptotic effects of theaflavin-3-gallate in human non-small cell lung cancer cells (A-549) along with determining the effect on cell cycle phase distribution, cell migration and invasion. Cell viability was determined by MTT assay while as phase contrast and fluorescence microscopies were involved to study apoptotic morphologi-cal features in these cells. Flow cytometry investigated the effect of theaflavin-3-gallate on cell cycle phase distribution. Theaflavin-3-gallate treatment led to a substantial cytotoxic effect in A-549 cancer cells with IC50 values of 42.1 µM and 27.9 µM at 24 and 48 hours respectively. Further, 80 and 160 µM dose of theaflavin-3-gallate induced apoptotic features including chromatin margina-tion and micronuclei presence. The population of cells in G2/M phase increased from 2.7% (control) to 6.8%, 17.2% and finally to 46.5% after treatment with 20, 80 and 160 µM concentration of theaflavin-3-gallate respectively indicating G2/M phase cell cycle arrest.

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Protein convertase subtilisin/Kexin type 9 inhibits hepatocellular carcinoma growth by interacting with GSTP1 and suppressing the JNK signaling pathway

10.21203/rs.3.rs-32718/v1 ◽

2020 ◽

Author(s):

Mingyan He ◽

Jing Hu ◽

Tingting Fang ◽

Wenqing Tang ◽

Bei Lv ◽

...

Keyword(s):

Cell Cycle ◽

Signaling Pathway ◽

Phase Distribution ◽

Cell Cycle Phase ◽

Cycle Phase ◽

Jnk Signaling ◽

Cell Cycle Phase Distribution ◽

Jnk Signaling Pathway

Abstract Background PCSK9 has been found to be closely related to the occurrence and development of a variety of tumors. However, the concrete role of PCSK9 and its relationship with HCC development is largely unknown. Methods The expression levels of PCSK9 in HCC tissues and HCC cell lines were determined by quantitative real-time polymerase chain reaction (qRT-PCR), western blot and immunohistochemistry assays. The effects of PCSK9 expression on HCC biological traits were investigated by overexpressing and downregulating PCSK9 protein in vivo and in vitro. The mechanism by which PCSK9 mediates the depolymerization of the GSTP1 dimer and the phosphorylation of the JNK signaling pathway was further investigated. Results PCSK9 is downregulated in human HCC tissues. HCC cell proliferation, cell cycle phase distribution and apoptosis were inhibited by PCSK9 in vitro. PCSK9 suppresses tumor growth and metastasis of HCC in vivo. PCSK9 interacts with GSTP1 and promotes the depolymerization of the GSTP1 dimer and inactivation of the JNK signaling pathway. Furthermore, low PCSK9 protein expression in primary HCC tissues correlated with worse clinical outcomes. Conclusions PCSK9 inhibits HCC cell proliferation, cell cycle phase distribution and apoptosis by interacting with GSTP1 and suppressing the JNK signaling pathway, suggesting that PCSK9 might act as a tumor suppressor and may serve as a therapeutic target for the treatment of HCC.

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