Vitamin D reverts resistance to the mTOR inhibitor Everolimus

Paper: Provvisiero DP et al. “Vitamin D reverts resistance to the mTOR inhibitor everolimus in hepatocellular carcinoma through the activation of a miR-375/oncogenes circuit.” Scientific Reports (2019) s41598-019-48081-9. (Nature)

1) Journal reputation & indexing

Peer review & scope. Scientific Reports (Nature Portfolio) is a large, peer-reviewed open-access “mega-journal” that judges manuscripts primarily on technical soundness rather than novelty. (Wikipedia)
Indexing. The journal is indexed in Web of Science, PubMed/MEDLINE, PubMed Central, Scopus, DOAJ, and others. (Nature)
Impact/metrics (context). 2024 Journal Impact Factor ≈ 3.9; 5-year IF ≈ 4.3. (Useful for context only; not a quality guarantee.) (Nature)

Takeaway: A reputable, widely indexed venue with rigorous but “soundness-focused” review criteria (not a selective, high-novelty journal). (Wikipedia)

2) Article content & methods

a) Study design

Model: strictly in-vitro HCC models; authors created everolimus-resistant (EveR) lines by chronic exposure (≥4 months) in PLC/PRF/5 and JHH-6; HepG2 EveR was generated but not used for the core analyses because resistance did not differ from parental cells. (Nature)
Interventions: calcitriol (1,25(OH)\₂D₃) at 10⁻⁷ M; everolimus at 10⁻¹⁴ to 10⁻⁸ M; vehicles kept at 0.04%. (Nature)
Assays: DNA content & colony formation for proliferation/survival; Western blot & immunofluorescence for EMT markers; qPCR (miRNA array and targeted); luciferase reporter to validate miR-375 binding to c-MYC (5′UTR); miR-375 mimic/inhibitor gain/loss-of-function. (Nature)

b) Validity of methods

The resistance model (long-term drug exposure) is standard for mechanistic studies. The miRNA causal chain(VitD → ↑miR-375 → ↓MTDH/YAP-1/c-MYC → resensitization) is probed with orthogonal tools (array screen → qPCR → luciferase → mimic/inhibitor), which strengthens internal validity. (Nature)
Limitations: entirely cell-based (no in-vivo data, no patient samples); everolimus concentrations are lower than typical clinical exposures (authors note this), which may limit translational relevance; WB experiments replicated twice, less robust than triplicate biological repeats used elsewhere in the paper. (Nature)

c) Data quality & statistics

Authors state quadruplicate experiments, replicated three times (except WB, twice). ANOVA with multiple-comparison corrections (Bonferroni/Newman-Keuls/Dunnett) via SPSS and GraphPad. This is acceptable for in-vitro work, though effect sizes and exact n’s per figure aren’t always explicit. (Nature)

d) Conclusions: objectivity & support

Main claim: calcitriol re-sensitizes everolimus-resistant HCC cells by up-regulating miR-375, which down-regulates MTDH, YAP-1, and c-MYC, and partly reverses EMT features. The data presented (functional assays + target validation) support this within the in-vitro context. Authors appropriately acknowledge that in-vitro only is a limitation and suggest future in-vivo work. (Nature)

3) References: relevance & currency

Citations span clinical HCC therapy, mTORi trials (including EVOLVE-1 everolimus failure), EMT/drug-resistance literature, and miRNA biology, with many sources from 2008–2018 (appropriate for a 2019 paper). (Nature)
Overall, the bibliography is technically relevant and reasonably current for the time of publication.

4) Author credibility & conflicts

Affiliations: University of Naples “Federico II,” Departments of Endocrinology and Public Health (academic lab setting). (Nature)
Funding: “This work was not supported by any grant.” (Neutral for bias; does imply resource limits.)
COI: “The authors declare no competing interests.”

5) How this paper should influence prior knowledge & its strength

What it adds: Mechanistic evidence that vitamin D (calcitriol) may overcome mTOR-inhibitor resistance in HCC via a miR-375–MTDH/YAP-1/c-MYC axis and partial EMT reversal. This extends prior observations that vitamin D and miRNAs modulate cancer phenotypes. (Nature)
Strength of influence: Hypothesis-generating / preclinical. Because the work is in-vitro only, with no animal efficacy or clinical correlation, it should modestly shift priors—encouraging in-vivo validation and biomarker-driven studies, but not changing clinical practice or guidelines by itself. Authors explicitly position it as a foundation for future in-vivo work. (Nature)

Bottom line

Scientific Reports is a peer-reviewed, widely indexed journal; the article meets standard technical-soundnesscriteria. (Nature, Wikipedia)
The experimental logic is coherent and multifaceted; statistics are standard for cell studies; COI absent. (Nature)
Key limitations—entirely in-vitro design, some modest replication depth, and use of sub-clinical drug concentrations—mean the findings should inform mechanism and future preclinical/clinical hypotheses, rather than directly influence clinical care. (Nature)

If you’d like, I can tabulate the paper’s risk-of-bias items (model selection, blinding, replication, reagent validation, statistics) and compare them with ARRIVE/NIH rigor guidelines to give you a scorecard-style summary.

Biohackerofthegods · September 9, 2025, 5:17pm

Wich is why i included so many of the studies and not just one

RapAdmin · September 9, 2025, 6:08pm

Have you read these studies / papers already? Why did you find them compelling? What do you think is the important message you’ve found in reading these papers? Please don’t just post a bunch of links to papers. There are millions of papers in Pubmed and we have no interest in people just copying them here into our forum without good reason. Your reasoning and rationale is not at all clear.

Biohackerofthegods · May 28, 2026, 1:33pm

This was suppose to be a PM

remove

Edit again: can post the vitamin d reply of my pm

Cause vitamin D is a mTOR inhibbitor, much better then rapamycin as it dosent supress it to hard so cell growth and repair stops, but instead putting the break on in an intellegent way while at the same time increase cell growth in tissue that needs it

Run it thrue AI and see for yourself

Btw have you seen that glp1s are the strongest senolytic drugs that ever been discovered? 1400% tumor supressive effect of reta, all of them induce autophagy

desertshores · May 28, 2026, 5:53pm

Damn, still another rabbit hole to explore. How did I miss that? What is the amount of mTORC1 suppression caused by vitamin D? I am only looking at D3 because that is what I take. The most recent research suggests we are not getting the optimal amount of vitamin D3.

So, what is the effect on mTORC1 of vitamin D3 while taking rapamycin? How do I calculate the effect of vitamin D3 and rapamycin on mTORC1? Apparently vitamin D3 has no effect on mTORC2.

Aristotle:

Conclusions

Evidence indicates that 1,25-dihydroxyvitamin D3 is a potent, selective inhibitor of mTORC1 signaling at concentrations between 1 nM and 1000 nM. This inhibition is achieved through the VDR-mediated induction of DDIT4/REDD1, which sequesters 14-3-3 proteins to activate the TSC1/TSC2 complex. Because mTORC1 disassembly is TSC-dependent while mTORC2 disassembly is not, Vitamin D selectively targets mTORC1 to modulate mitochondrial metabolism, upregulate glucocorticoid receptors, and induce autophagy. Future research must address the critical gap in quantitative pharmacology by establishing precise IC50 values and directly measuring mTORC2 substrates under controlled Vitamin D titration.

Full response here:

Review what is known about vitamin D and inhibition of mTORC1 vs. mTORC2.

So, I was wondering how many nM 10,000 vitamin D3 taken daily would produce in the blood of a 70 kg adult. And I did find a calculator. Vitamin D3 Dosage Calculator