Arylquin 1 (Potent Par-4 Secretagogue) Inhibits Tumor Progression and Induces Apoptosis in Colon Cancer Cells

Round 1

Reviewer 1 Report

The authors aimed to investigate the role of Arylquin 1  as an inducer of apoptosis in colorectal cancer cells. the topic is little studied, and there are no articles in the literature.

However, Arylquin 1, is regarded as a secretagogue of prostate apoptosis response-4 (Par-4), of which there are many articles in the literature.
The authors must enrich the introduction, which is too brief, by adding the various studies concerning Par-4 and its role in colorectal carcinogenesis.

The discussion needs to be broadened and deepened, the experimental results are not clearly discussed.
For example, regarding the MAPK signaling pathway, its role and importance in colorectal carcinogenesis must be investigated.
The authors further state that Arylquin 1 attenuates EMT in CRC, the sentence must be detailed and explained.

Author Response

Please see the attachment.

Thank you.

Author Response File: Author Response.docx

Reviewer 2 Report

The authors present preliminary results regarding the effects of Arylquin 1 in SW620 and HCT116 cell lines. The manuscript is well organized but, from my point of view, minor and major revisions must be done before acceptance.

Major revisions

The introduction briefly describes Arylquin 1 and its effects on some tumor types. I suggest enriching the introduction regarding CRC treatment and explain / highlight the importance of a new drug therapy.

In Figure 3 you show the figures and blots with different Arylquin 1 concentrations in a specific time after administration. In the legend you say 0, 16, 24, and 48 hours after treatment but this is not shown in the figure. Please add the figures of the different incubation time to compare them. In the text (lines 73 -76) you only mention the 24hs treatment. Do the other administration time (16hs and 48hs) show significant cell invasion inhibition?

At section 2.1 you obtained IC50 concentrations of 1.8 and 2.3 µM for SW620 and HCT116, respectively. For the following experiments you test 1-, 1.5-, and 2- μM for both cell lines. Since 2.3 μM is the IC50 concentration for HCT116, can you explain why you choose lower concentrations. The selected concentrations exclude 2.3 μM and so you may lose a significant effect on this cell line upon treatment.

Please add the hours after treatment in section 2.3 and Figure 5.

As shown in section 2.3, results show a different behaviour of SW620 and HCT116. HCT116 presents a significant decrease in BCL-2 expression and higher apoptosis percentage than SW620. I suggest explaining cell line differences in the discussion.

Discussion: The statement you write in line 174-177 is not correct: BCL2 is not downregulated in SW620.

Discussion (line 183-197): in the last paragraph of the discussion you discuss the results of section 2.4. You start with the following statement: “In the present study, we found decreased expression of phosphorylated ERK in CRC 183 treated with Arylquin 1, in addition to JNK and p38 downregulation”. In the blots of Figure 7 it is observed a constant and significant increase upon treatment. The last paragraph of the discussion contradicts the results of section 2.4. ERK, p38 and JNK are mainly involved in tumor progression. Their upregulation observed in Figure 7 after treatment does not support the anti-tumoral role of Arylquin 1.

Minor revisions

In line 46: use past participle after to be. You write: “…have yet to be determinate”. Please change determinate to determinated.

In Figure 6 I suggest moving the percentage of viable cells, so it’s clear.

Why do you choose HCT116 for tumor xenograft? Please specify

Author Response

Please see the attachment.

Thank you.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

accept in the present form

Reviewer 2 Report

The authors clearly answer to my questions and suggestions. The revised manuscript improves the presentation quality and the scientific soundness of the research work.

I point out a minor revision:

In line 218 you write: "… due to promote its localization in cell surface". Maybe you want to say " ... due to its localization in cell surface".