Chemopreventive effects of vitamin D3 and its analogue, paricalcitol, in combination with 5-fluorouracil against colorectal cancer: The role of calcium signalling molecules

Aslam, Akhmed; Ahmad, Jawwad; Baghdadi, Mohammed; Idris, Shakir; Almaimani, Riyad A.; Alsaegh, Aiman; Alhadrami, Mai; Refaat, Bassem

doi:10.1016/j.bbadis.2020.166040

Cited by 18 publications

(27 citation statements)

References 58 publications

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“…The aforementioned clinical trials verify that vitamin D 3 inhibits colorectal carcinogenesis and increases sensitivity to chemotherapy. Furthermore, numerous studies have successively come to similar conclusions by in vitro and in vivo experiments (Padi et al, 2013;Razak et al, 2019;Shang et al, 2020;Aslam et al, 2021). A notable study in pancreatic cancer showed that vitamin D 3 increased gemcitabine concentration in tumor tissues; thus, tumors were smaller and animals survived longer than when gemcitabine alone was given (Sherman Mara et al, 2014).…”

Section: Discussionmentioning

confidence: 85%

VDR Signaling via the Enzyme NAT2 Inhibits Colorectal Cancer Progression

et al. 2021

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Recent epidemiological and preclinical evidence indicates that vitamin D3 inhibits colorectal cancer (CRC) progression, but the mechanism has not been completely elucidated. This study was designed to determine the protective effects of vitamin D3 and identify crucial targets and regulatory mechanisms in CRC. First, we confirmed that 1,25(OH)2D3, the active form of vitamin D3, suppressed the aggressive phenotype of CRC in vitro and in vivo. Based on a network pharmacological analysis, N-acetyltransferase 2 (NAT2) was identified as a potential target of vitamin D3 against CRC. Clinical data of CRC patients from our hospital and bioinformatics analysis by online databases indicated that NAT2 was downregulated in CRC specimens and that the lower expression of NAT2 was correlated with a higher metastasis risk and lower survival rate of CRC patients. Furthermore, we found that NAT2 suppressed the proliferation and migration capacity of CRC cells, and the JAK1/STAT3 signaling pathway might be the underlying mechanism. Moreover, Western blot and immunofluorescence staining assays demonstrated that 1,25(OH)2D3 promoted NAT2 expression, and the chromatin immunoprecipitation assay indicated that the vitamin D receptor (VDR) transcriptionally regulated NAT2. These findings expand the potential uses of vitamin D3 against CRC and introduce VDR signaling via the enzyme NAT2 as a potential diagnostic and therapeutic target for CRC.

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Section: Discussionmentioning

confidence: 85%

VDR Signaling via the Enzyme NAT2 Inhibits Colorectal Cancer Progression

et al. 2021

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“…Following treatment with activins or follistatin, the cells were processed for cell cycle analysis according to Aslam et al [ 26 ]. Briefly, cells were trypsinised, washed twice with PBS (500× g for 5 min), and then fixed in ice-cold 70% ethanol for 24 h at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Profiling Activins and Follistatin in Colorectal Cancer According to Clinical Stage, Tumour Sidedness and Smad4 Status

Refaat¹,

Zekri²,

Aslam³

et al. 2021

Pathol. Oncol. Res.

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This study explored the roles of activins and follistatin in colorectal cancers. Paired malignant and normal colonic tissues were collected from archived paraffin-embedded (n = 90 patients) alongside fresh (n = 40 patients) specimen cohorts. Activin β-subunits, follistatin and Smad4 mRNAs and proteins were measured by real-time PCR and immunohistochemistry (IHC). Mature activin-A, -B, -AB and follistatin proteins were measured by ELISA. Cancer tissues having ≤ the 20th percentile of the Smad4 IHC score were considered as low (L-S4) group. The Smad4-intact SW480 and Smad4-null HT29 colon cancer cell lines were treated with activins and follistatin, and cell cycle was analysed by flow cytometry. The cell cycle inducing (CCND1/CCND3) and inhibitory (p21/p27) proteins alongside the survival (survivin/BCL2) and pro-apoptosis (Casp-8/Casp-3) markers were measured by immunofluorescence. Thirty-nine patients had right-sided cancers (30%) and showed higher rates of L-S4 tumours (n = 17; 13.1%) alongside worse clinicopathological characteristics relative to left-sided cancers. The βA-subunit and activin-A increased, whilst βB-subunit and activin-AB decreased, in malignant sites and the late-stage cancers revealed the greatest abnormalities. Interestingly, follistatin declined markedly in early-stage malignant tissues, whilst increased significantly in the advanced stages. All activin molecules were comparable between the early stage right- and left-sided tumours, whereas the late-stage right-sided cancers and L-S4 tumours showed more profound deregulations. In vitro, activin-A increased the numbers of the SW480 cells in sub-G1 and G0/G1-phases, whereas reduced the HT29 cell numbers in the sub-G1 phase with simultaneous increases in the G0/G1 and S phases. The p21/p27/Casp-8/Casp-3 proteins escalated, whilst CCND1/CCND3/BCL2/survivin declined in the SW480 cells following activin-A, whereas activin-A only promoted p21 and p27 alongside reduced CCND3 in the HT29 cells. By contrast, activin-AB increased the numbers of SW480 and HT29 cells in Sub-G1 and G0/G1-phases and promoted the anti-cancer and reduced the oncogenic proteins in both cell lines. In conclusion, activins and follistatin displayed stage-dependent dysregulations and were markedly altered during the advanced stages of right-sided and L-S4 cancers. Moreover, the activin-A actions in CRC could be Smad4-dependent, whereas activin-AB may act as a Smad4-independent tumour suppressor protein.

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“…Freshly prepared calcitriol (0.07 µg/Kg/day; ve times/week) and metformin (430 mg/Kg/day; ve times/week) were administrated orally to the designated groups, and the delivered amounts were equivalent to the highest daily recommended doses for a 60 Kg body weight adult human for calcitriol (0.25 µg/day; 0.0042 µg/Kg/day) [34] and metformin (1500 mg/day; 25 mg/Kg/day) [35] as per the dose conversion formula between human and mice [36]. 5-FU was administrated to the designated groups for four successive cycles as a single weekly intraperitoneal injection (50 mg/Kg/week) as previously described [21].…”

Section: Experimental Animal Studies and Treatment Protocolsmentioning

confidence: 99%

“…In this regard, several studies revealed negative correlations between the levels of serum vitamin D and CRC development [18,19]. Moreover, active vitamin D 3 (VD 3 ) attenuated the expression of the PI3K/Akt/mTOR pathway, inhibited cell proliferation and promoted apoptosis in colon cancer cell lines [20][21][22][23]. Other studies have also reported potent anti-tumorigenic effects for metformin against CRC that were portrayed by cell cycle arrest, increased apoptosis, and downregulations in the PI3K/Akt/mTOR network [24][25][26][27][28].…”

Section: Introductionmentioning

confidence: 99%

“…Other studies have also reported potent anti-tumorigenic effects for metformin against CRC that were portrayed by cell cycle arrest, increased apoptosis, and downregulations in the PI3K/Akt/mTOR network [24][25][26][27][28]. Additionally, others have shown enhanced tumoricidal actions against CRC by combining VD 3 or metformin with 5-FU [21,[29][30][31], as well as by adding both agents together [32,33].…”

Section: Introductionmentioning

confidence: 99%

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Metformin and Calcitriol Enhance 5-Fluorouracil Tumoricidal Effects in Colon Cancer By Modulating The PI3K/Akt/PTEN/mTOR Network

Almaimani

Aslam

Ahmad

et al. 2022

Preprint

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Purpose: Chemoresistance to 5-Fluorouracil (5-FU) is common during colorectal cancer (CRC) treatment. This study measured the chemotherapeutic effects of 5-FU, calcitriol, and/or metformin single/dual/triple regimens as complementary/alternative therapies. Methods: Ninety male mice were divided into: negative and positive (PC) controls, 5-FU, Cal, Met, 5-FU/Cal, 5-FU/Met, Cal/Met, and 5-FU/Cal/Met groups. Treatments lasted four weeks following CRC induction by azoxymethane. The therapeutic regimens were also applied in the SW480 and SW620 CRC cell lines. Results: The PC mice had abundant tumours, markedly elevated proliferation markers (survivin/CCND1) and PI3K/Akt/mTOR alongside reduced p21/PTEN/Cytochrome-C/Caspase-3 and apoptosis. All therapies reduced tumour numbers, with 5-FU/Cal/Met most prominent regimen. All protocols also decreased cell proliferation markers, inhibited PI3K/Akt/mTOR molecules, increased pro-apoptotic molecules with apoptosis index, and 5-FU/Cal/Met revealed the strongest anti-cancer effects. In vitro, all therapies equally induced G1-phase arrest in SW480 cells, whereas metformin-alone showed maximal SW620 cell numbers in G0/G1-phase. 5-FU/Met co-therapy also showed the highest apoptotic SW480 cell numbers (13%), whilst 5-FU/Cal/Met disclosed the lowest percentage (81%) of viable SW620 cells. Moreover, 5-FU/Cal/Met revealed maximal inhibitions of cell cycle inducers (CCND1/CCND3), cell survival (BCL2) and the PI3K/Akt/mTOR molecules alongside highest expression of cell cycle inhibitors (p21/p27), pro-apoptotic markers (BAX/Cytochrome-C/Caspase-3), and PTEN in both cell lines. Conclusions: Metformin monotherapy was superior to calcitriol, whereas the 5-FU/metformin protocol showed better anti-cancer effects relative to the other dual therapies. However, the 5-FU/Cal/Met approach displayed the best in vivo and in vitro tumoricidal effects related to cell cycle arrest and apoptosis, justifiably by enhanced modulations of the PI3K/PTEN/Akt/mTOR pathway.

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Chemopreventive effects of vitamin D3 and its analogue, paricalcitol, in combination with 5-fluorouracil against colorectal cancer: The role of calcium signalling molecules

Cited by 18 publications

References 58 publications

VDR Signaling via the Enzyme NAT2 Inhibits Colorectal Cancer Progression

VDR Signaling via the Enzyme NAT2 Inhibits Colorectal Cancer Progression

Profiling Activins and Follistatin in Colorectal Cancer According to Clinical Stage, Tumour Sidedness and Smad4 Status

Metformin and Calcitriol Enhance 5-Fluorouracil Tumoricidal Effects in Colon Cancer By Modulating The PI3K/Akt/PTEN/mTOR Network

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