Anti-Metastatic Activity of Tagitinin C from Tithonia diversifolia in a Xenograft Mouse Model of Hepatocellular Carcinoma

Round 1

Reviewer 1 Report

Dear the Editor

Lin C-Y et al described the anti-metastatic activity of tagitinin C in the mouse model od hepatocellular carcinoma. First, using methanolic extract of Tithonia difersifolia leaves, these authors performed an in vitro study. Then, they found an impaired tumorigenicity of Hep-G2 and Huh 7 cell xenograft in mouse model. This study was well-planned and organized in sound.

 

Major concerns: 

1) Some audience might be interested in the mode of action of taginin C, a sesquiterpenoid in this plant acting as an inhibitor for cell proliferation. Any discussion on this point would be appreciated.

2) If possible, please discuss the purity of this preparation. Is there any chromatographic data?

 

Minor comments:

1) In Table 1, for which parameters for IC50 vaules for tagnitin C was presented? Is this for cell viability? A unit for IC50 needs to be indicated. Please also explain the meaning of * in clone 9.

2) In Figure 3, a scale bar needs to be inserted.

3) In Table 3, units for each parameter needs to be clearly indicated.

Author Response

Dear reviewer,

I am very grateful for your comments about the manuscript. According with your advice, we amended the relevant part in manuscript: table 1, table 3, figure 3, and line 83-85, 265-271. The star symbol in table 1 is typo, and we have already modified it. The HPLC analysis certified the content of tagitinin C (as attached file), the major sesquiterpenoid in theTithonia diversifolia methanolic extract (TDM), as 1.45 % and the purity is more than 95 %. We hope that meets your requirement.

Sincerely

Author Response File: Author Response.docx

Reviewer 2 Report

In this paper, Lin et al. describe the anti-metastatic activity of tagitinin C from Tithonia diversifolia in in vitro studies and in xenograft mouse model of hepatocellular carcinoma.

General comment: The results of the study are relevant and interesting.

Specific points:

Figure 1: Could the resolution be improved?

Figure 3: Could the scratch assay images be enlarged?

The authors say: 'Untreated Hep-G2 or Huh 7 cells (1.5 × 107) were injected subcutaneously into a single site of each mouse'. Could the authors specify in which site the cells were injected?

The authors say: 'Xenograft models of human HCC exhibit diffuse compact trabeculae with variable degrees of anaplasia having increased mitotic and metastatic activity and irregular masses. Hence, tumor volume and solid mass are not suitable anti-tumorigenic indicators, as reported in other studies [12].' Since metastasis are correlated with the primary tumors (volume and solid mass), could the authors elaborate on how this fact is considered in their study?

The authors say: 'First, we anesthetized the rats using sodium pentobarbital…'. Is this a mistake, since the mice were used in the study?

Author Response

Dear reviewer,

I am very grateful for your comments about the manuscript. According with your advice, we amended the relevant part in manuscript: figure 1, figure 3, and line: 62-63. Hep-G2 or Huh 7 cells were injected into the dorsum of each mouse. The “rats” is typo, and we have modified it. The content of reference has been rechecked, and we have amended the relevant part in manuscript.

Sincerely

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

Dear the Editor

The revised manuscript properly addressed the raised concerns by this Reviewer.