Raghavan, J Cancer Sci Ther 2018, 10:11
DOI: 10.4172/1948-5956.1000567
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Cancer Science & Therapy
ISSN: 1948-5956
Research Article
Open Access
Metadichol® a Novel Agonist of the Anti-aging Klotho Gene in Cancer Cell
Lines
Palayakotai R Raghavan*
Nanorx Inc., PO Box 131, Chappaqua, NY 10514, USA
Abstract
Klotho is an anti-aging protein that is mostly secreted by the kidneys, the brain, and the thyroid. It plays a
significant role in regulating kidney function and vascular health. Klotho gene is named after "the Spinner" (Clotho
from Greek mythology), the goddess who spins the thread of life. Klotho is a transmembrane protein known to be a
co-receptor for Fibroblast Growth Factor-23. Klotho gene is expressed in a variety of tissues changes in the levels
are associated with many diseases. Klotho is a tumor suppressor in breast cancer and its expression is reduced
in human pancreatic adenocarcinoma, and treatment with klotho inhibits the growth of pancreatic cancer cells in
vitro and in vivo.
Growing evidence suggests that an increase in KL expression may be beneficial for age-related diseases
such as arteriosclerosis and diabetes. It remains a challenge today to induce Klotho expression. Herein we show
that treating pancreatic cancer cells PANC1, MIAPACA and COLO-205 with Metadichol® a novel food based lipid
emulsion of long chain alcohols at picogram/ml, concentration led to a 4-10 fold increase in Klotho expression as
seen quantitative RT-PCR. These results suggest the use of Metadichol® given its constituents that are present in
foods we consume every day is a novel therapeutic intervention for pancreatic cancer and other diseases.
Keywords: VDR; Metadichol; Klotho; Inverse agonist; Protean
agonist; Constitutive receptors; Pancreatic cancer; FGF-23; Diabetes;
Anti-aging; PANC1; COLO-205; MIAPACA; Long chain alcohols
Introduction
The Greek goddess whose name is associated with Klotho protein
spins life’s thread and is associated with reversing aging in mammals.
In Greek Mythology, Klotho has two siblings, Lachesis and Atropos,
and one determines the length of the thread of life and the other cuts the
thread. Klotho (KL), which was named after one of the three goddesses
of fate who controlled aging in Greek mythology, was initially identified
in 1997 as the gene responsible for early aging-like symptoms in mice
[1], and in several other tissues [2]. It acts as a coreceptor with fibroblast
growth factor receptor-1 (FGFR1)to bind fibroblast growth factor 23
(FGF23) and mediate phosphaturia to correct the hyperphosphatemia
arising from 1,25-dihydroxy vitamin D (calcitriol or 1,25D) Stimulation
of intestinal calcium and phosphate absorption.1,25D regulates the
expression of both membrane and soluble klotho forms in multiple kidney
cell types to support FGF23 phosphaturic and vitamin D counter-regulatory
actions at the kidney, possibly exerting antiaging effects [3].
The observation that Klotho inhibits insulin/IGF-1 signalling has
ramifications for therapeutic intervention in cancer as well. Activation
of the IGF receptor has been implicated in the etiology of carcinomas
[4]. There is a growing body of evidence implicating Klotho as a tumour
suppressor [5]. In particular cervical, colorectal, gastric and lung
carcinoma, pancreatic, hepatocellular carcinoma and breast cancer
amongst a few [6]. In general, Higher Klotho expression was associated
with smaller tumor size and Klotho treatment slowed the progression
of cancer. Klotho is significantly down regulated in all cancer types
including brain malignancies [7]. Down regulation of Klotho (Table
1) was observed across the different cancer types. The role of Klotho
in cancer as a tumor suppressor mentioned by Wolf I [8] showed how
Klotho putative tumor suppressor in breast cancer.
The available data indicate that Klotho acts as a universal tumor
suppressor and that there may be a role for Klotho cancer treatment.
Currently, there are no Klotho-based treatments available, although a
J Cancer Sci Ther, an open access journal
ISSN: 1948-5956
number of commonly used compounds do either directly up-regulate
Klotho in vitro, like PPARγ agonists [9], vitamin D [10], Testosterone
[11] and Resveratrol [12], or otherwise up-regulate or at least inhibit
down-regulation of Klotho in vivo. Recent data indicate that Klotho
has extensive effects over the entire spectrum of human diseases [13]
as shown in Table 1.
Acetylcholine and Nitric Oxide
Dysregulation Aging (highly
accelerated)
Bone Loss (such as osteoporosis
and low bone mass)
All-cause mortality
Cancer
Anemia
Cataracts
Anorexia
Chronic stress
Atherosclerosis (as well as calcification of
the arteries)
Depression
Growth hormone deficiency
Diabetes
Kidney disease (such as CKD and
electrolyte imbalances) Kidney transplant
Glaucoma
Hyperphosphatemia
Multiple system atrophy
Hyperparathyroidism
Pseudoexfoliation syndrome
Hypertension
Rheumatoid arthritis
Impaired cognition (such as Alzheimer's
Disease)
Sarcopenia
Inflammatory bowel disease
Skin atrophy (such as scleroderma)
Lung damage
Vascular disease (such as coronary
artery disease)
Stroke
--
Table 1: Klotho effects over the entire spectrum of human diseases
*Corresponding author: Palayakotai R Raghavan, Nanorx Inc., PO Box 131,
Chappaqua, NY 10514, USA, Tel: +1-914-671-0224; E-mail: raghavan@nanorxinc.com
Received October 11, 2018; Accepted November 08, 2018; Published November
10, 2018
Citation: Raghavan PR (2018) Metadichol® a Novel Agonist of the Anti-aging Klotho
Gene in Cancer Cell Lines. J Cancer Sci Ther 10: 351-357. doi: 10.4172/19485956.1000567
Copyright: © 2018 Raghavan PR. This is an open-access article distributed under
the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and
source are credited.
Volume 10(11) 351-359 (2018) - 351
Citation: Raghavan PR (2018) Metadichol® a Novel Agonist of the Anti-aging Klotho Gene in Cancer Cell Lines. J Cancer Sci Ther 10: 351-357. doi:
10.4172/1948-5956.1000567
Klotho levels are decreased in certain types of human tumor cells/
tissues, and reduced levels are associated with decreased survival [14].
Animal studies show that Klotho can improve survival rates [15] reduce
metastasis and reduce cancer cell resistance to chemotherapeutic agents
[16]. Abramovitz [17] showed in studies on pancreatic adenocarcinoma
cell lines that Klotho expression is reduced, and treatment with Klotho
effectively slows growth of pancreatic cancer cells in vitro and in
vivo. Biao Xie also have shown that Klotho is a tumor suppressor in
gastric cancer [18]. Injection of secreted Klotho protein suppressed
metastasis and improved survival in mice transplanted with human
lung cancer cells [19]. Long-term administration of KL to mice shows
a favorable toxicity profile. As klotho is an endogenous hormone, its
administration is potentially feasible and may serve as a novel therapy
for pancreatic, as well as other cancers.
Metadichol® nano-emulsion of long chain alcohols is an inverse
agonist of VDR (Vitamin D receptor) that is non- toxic. We tested it in
the pancreatic cell lines PANC1, COLO-205 and MIAPACA cell lines
and the results show that it enhances Klotho expression and thus would
pave the way for use a therapeutic in diseases where increased Klotho
levels are required.
Experimental
The experimental work was outsourced and carried out by Skanda
Life Sciences Private Limited of Bangalore India. The cell lines were
purchased from ATCC, USA and primers from Eurofins India. PCR
and qPCR Method Standardization. For each of target gene the PCR
conditions viz, Tm, Amplicon specificity & size were optimized using
in-house established and validated methods/reagents.
qPCR
Instruments used CFX96 real time PCR, Bio-Rad. Gene regulation
of KLOTHO genes in MiaPaca, Colo-205, Panc-1 cells treated with
metadichol.
Cell lines
COLO 205 (ATCC® CCL-222™), MIA PaCa-2 (ATCC® CRL-1420™),
PANC-1 (ATCC® CRL-1469™) (Table 2).
Sample Preparation and RNA Isolation
Total RNA from the cells was extracted using TRizol Reagent
(Invitrogen) according to manufacturer’s instruction. Cells were
washed twice with PBS and centrifuged at 2000rpm for 5min. To the
cell pellet, 1ml of TRIzol (per p35 dish) was added in 1.5ml Eppendrof
Sample
tube and vortexed. Samples were allowed to stand for 5 minutes at room
temperature. To the reaction mixture 0.2 ml of chloroform is added
and vigorously mixed for 15 seconds. The tube was allowed to stand
at room temperature for 5 minutes, centrifuged the resulting mixture
at 10,000rpm for 15min at 40 C. Upper aqueous phase is transferred
to a new clean Eppendorf tube and treated with 0.5ml of isopropanol.
The resultant mixture is mixed gently by inverting the sample 5 times
and incubated at room temperature for 5 minutes. Samples were
centrifuged at 10,000 rpm for 10 minutes at 40 C. Supernatant liquid
was discarded and the RNA pellet was washed by adding 1ml of 70%
ethanol. Mix the sample gently by inverting few times. Centrifuged for
5min at 14,000rpm at 40 C. Supernatant was discarded by inverting the
tube on a clean tissue paper. Later, the pellet was dried by incubating in
a dry bath for 5min at 550 C. The pellet was then resuspended in 25 µl
of DEPC treated water.
RT-PCR
A semi quantitative reverse transcriptase polymerase chain reaction
(RT-PCR) was carried out using Techno Prime system to determine
the levels of Klotho and β-Actin mRNA expressions. The cDNA was
synthesized from 2 µg of RNA using the Verso cDNA synthesis kit
(Thermo Fischer Scientific) with oligo dT primer according to the
manufacturer’s instructions. The reaction volume was set to 20μl and
cDNA synthesis was performed at 42o C for 60 min, followed by RT
inactivation at 85o C for 5 min (Table 3).
PCR
The PCR mixture (final volume of 20 µL) contained 1 µL of
cDNA, 10 µL of Red Taq Master Mix 2x (Amplicon) and 1µM of
each complementary primer specific for Klotho and β-Actin (internal
control) sequence. The samples were denatured at 94o C for 5 minutes
and amplified using 35 cycles of 94o C for 30 seconds, 53o C for 30
seconds, and 72o C for 1 minute for KLOTHO renaturation was
set to 49o C and for β-Actin the renaturation was set to 55o C for 30
seconds followed by a final elongation at 72o C for 10 minutes. The
optimal numbers of cycles have been selected for amplification of this
genes experimentally so that amplifications were in the exponential
range and had not reached a plateau. Ten microliters of the final
amplification product were run on a 2% ethidium-stained agarose gel
and photographed. Quantification of the results was accomplished by
measuring the optical density of the bands, using the computerized
imaging program Image J. The values were normalized to β-Actin
intensity levels (Figure 1-10).
Cell culture condition
Treatment
Control (Media)
1 pg/ml
100 pg/ml
MIAPACA COLO-205, PANC-1 cells
(1 × 106) grown in P35 dish were treated with test compound
Metadichol
1 ng/ml
100 ng/ml
1 µg/ml
Table 2: Treatment protocol.
Gene
Primer pair
Sequence
Tm
Β-Actin
FP
TCCTCCTGAGCGCAAGTACTCT
62.1
RP
GCTCAGTAACAGTCCGCCTAGAA
62.4
FP
GGGAGGTCAGGTGTCCATTG
55.88
RP
TGCTCTCGGGATAGTCACCA
53.83
Klotho
Product size (bp)
153
152
Table 3: Primer details.
J Cancer Sci Ther, an open access journal
ISSN: 1948-5956
Volume 10(11) 351-359 (2018) - 352
Citation: Raghavan PR (2018) Metadichol® a Novel Agonist of the Anti-aging Klotho Gene in Cancer Cell Lines. J Cancer Sci Ther 10: 351-357. doi:
10.4172/1948-5956.1000567
Figure 1: Amplification of β-Actin gene in MIAPACA (Lane 1-Ladder; Lane 2-Control; Lane 3-1 pg/mL; Lane 4-100 pg/mL; Lane 5-1 ng/ml; Lane 6-100 ng/ml;
Lane 7-1 µg/mL).
Figure 2: Amplification of Klotho gene in MIAPACA cell (Lane 1-Ladder; Lane 2-Control; Lane 3-1 pg/mL; Lane 4-100 pg/mL; Lane 5-1 ng/ml; Lane 6-100 ng/
ml; Lane 7-1 µg/mL).
Figure 3: Amplification of β-Actin gene in COLO-205 (Lane 1- Ladder; Lane 2-Control; Lane 3-1 ng/mL; Lane 4-100 ng/mL; Lane 5-1 pg/ml; Lane 6-100 pg/ml;
Lane 7-1 µg/mL).
J Cancer Sci Ther, an open access journal
ISSN: 1948-5956
Volume 10(11) 351-359 (2018) - 353
Citation: Raghavan PR (2018) Metadichol® a Novel Agonist of the Anti-aging Klotho Gene in Cancer Cell Lines. J Cancer Sci Ther 10: 351-357. doi:
10.4172/1948-5956.1000567
Figure 4: Amplification of Klotho gene in COLO-205 cell (Lane 1- Ladder; Lane 2-Control; Lane 3-1 ng/mL; Lane 4-100 ng/mL; Lane 5-1 pg/ml; Lane 6-100 pg/
ml; Lane 7-1 µg/mL).
100 bp
100 bp
Figure 5: Amplification of β-Actin gene in PANC1 (Lane 1- Ladder; Lane 2-Control; Lane 3-1 ng/mL; Lane 4-100 ng/mL; Lane 5-1 pg/ml; Lane 6-100 pg/ml; Lane
7-1 µg/mL).
Figure 6: Amplification of Klotho gene in PANC1 cell (Lane 1- Ladder; Lane 2-Control; Lane 3-1 ng/mL; Lane 4-100 ng/mL ; Lane 5-1 pg/ml; Lane 6-100 pg/ml;
Lane 7-1 µg/mL).
J Cancer Sci Ther, an open access journal
ISSN: 1948-5956
Volume 10(11) 351-359 (2018) - 354
Citation: Raghavan PR (2018) Metadichol® a Novel Agonist of the Anti-aging Klotho Gene in Cancer Cell Lines. J Cancer Sci Ther 10: 351-357. doi:
10.4172/1948-5956.1000567
Figure 7: Raw data of MIAPACA cell line.
J Cancer Sci Ther, an open access journal
ISSN: 1948-5956
Volume 10(11) 351-359 (2018) - 355
Citation: Raghavan PR (2018) Metadichol® a Novel Agonist of the Anti-aging Klotho Gene in Cancer Cell Lines. J Cancer Sci Ther 10: 351-357. doi:
10.4172/1948-5956.1000567
Figure 8: Raw data COLO-205 cell line.
Results
Three different cell lines were individually treated with Metadichol
at various concentrations to assess the expression of Klotho gene. The
maximum up regulation of Klotho gene expression is seen at lowest
concentration treated, i.e., 1pg/ml in both MIAPACA and PANC-1
J Cancer Sci Ther, an open access journal
ISSN: 1948-5956
cells up 10.34 and 3.66-fold, whereas, in COLO 205 the expression at
1µg/ml was maximum up 6.36-fold compared to control. Overall, the
Up regulation of Klotho gene expression level is dose dependent in
MIAPACA cells from highest to lowest treatment concentrations from
1µg/ml to 1pg/ml.
Volume 10(11) 351-359 (2018) - 356
Citation: Raghavan PR (2018) Metadichol® a Novel Agonist of the Anti-aging Klotho Gene in Cancer Cell Lines. J Cancer Sci Ther 10: 351-357. doi:
10.4172/1948-5956.1000567
Figure 9: Raw data PANC1 cell line.
J Cancer Sci Ther, an open access journal
ISSN: 1948-5956
Volume 10(11) 351-359 (2018) - 357
Citation: Raghavan PR (2018) Metadichol® a Novel Agonist of the Anti-aging Klotho Gene in Cancer Cell Lines. J Cancer Sci Ther 10: 351-357. doi:
10.4172/1948-5956.1000567
Figure 10: Summary of results Q-RT-PCR of Meta-dichol treated cells.
Discussion
King have identified small molecules that elevated Klotho
expression, but the increase was only in the range of 20-50% at micro
molar (uM) concentrations and with compounds whose toxicity is not
known in humans [20].
Marco [21] suggested that Orally-available, transcriptional factors
like D-alpha-tocopherol [22], and vitamin D receptors (VDR) agonists
such as cholecalciferol [23,24] and lithocholic acid [25] can increase
Klotho expression. They suggested that, or a combination of these
molecules would result in increased expression of endogenous, human
Klotho through transcriptional activation.
R.E. Forster [26] have postulated that the liganded VDR up
regulates Klotho gene via Vitamin D response elements (VDRE). The
actions of 1,25 dihydroxyvitamin D3 (1,25 D3) on phosphorus are
opposed via the combined effects of FGF23 and Klotho, which is up
regulated by the liganded vitamin D receptor.
1,25D3 acting on VDR induces FGF23 in osteocytes to increase
circulating FGF23 [27], which protects against hyperphosphatemia
[28]. FGF23 also increases 1,25D3 degradation [29].
Metadichol® a nanoemulsion of long-chain binds to VDR Receptor
as an inverse agonist, and the formulation contains D-alpha-tocopherol
[30]. Inverse agonists bind to the same binding site as the agonists
in case of VDR it is 1,25 dihydroxy Vitamin D3. They induce a
pharmacological response different and distinct when compared to that
of the agonist. Metadichol in our human subjects [31] behaves more
likely a Protean agonist as it exhibits dual properties like, e.g. Increasing
Insulin Secretion (type 1) and reducing Insulin (type 2). Protean
agonists behave as both positive and negative agonists on the same
receptor, depending on the degree of constitutive activity. If there is no
constitutive activity, the agonist would be a positive agonist [32]. When
J Cancer Sci Ther, an open access journal
ISSN: 1948-5956
constitutive activity is present, the Protean agonist would be an inverse
agonist. Metadichol an extract of sugar cane wax exhibits properties
that could also be considered as an Adaptogens [33] which are unique
in their ability to balance endocrine hormones and the immune system
[34-37]. Adaptogens help maintain optimal homeostasis in the body.
Adaptogens are proposed to have a normalizing in the body effect
on the body and have the ability to toning down the activity of hyper
functioning systems in case of constitutive receptors or strengthening
the operation of hypo-functioning systems like an agonist. Given the
precedence of VDR and its role in regulating Klotho genes it is not
surprising that Metadichol® actions on VDR have a similar outcome
and as a hormone, it shows activity at picogram levels. Also, Metadichol
is nontoxic [38-40] as compared to other solutions in literature to
enhance the use of Klotho as a therapeutic target.
Conclusion
Klotho has been shown to have a wide range of roles in many
pathologies. Changes in the levels of Klotho are associated with many
diseases. It could be useful as a potential biomarker. However, also
has a future as a safe therapeutic in mitigating various diseases where
Klotho has a significant role (41).
References
1. Kuro-o M, Matsumura Y, Aizawa H, Kawaguchi H, Suga T, et al. (1997)
Mutation of the mouse klotho gene leads to a syndrome resembling aging.
Nature 390: 45-51.
2. Kuro-o M (2010) Klotho. Pflugers Arch 459: 333-343.
3. Haussler MR, Haussler CA, Whitfield GK, Hsieh JC, Thompson PD, et
al. (2010) The nuclear vitamin D receptor controls the expression of genes
encoding factors which feed the ‘‘Fountain of Youth’’ to mediate healthy aging.
J Steroid Biochem Mol Biol 121: 88-97.
4. Pollak MN, Schernhammer ES, Hankinson SE (2004) Insulin-like growth
factors and neoplasia. Nat Rev Cancer 4: 505-518.
Volume 10(11) 351-359 (2018) - 358
Citation: Raghavan PR (2018) Metadichol® a Novel Agonist of the Anti-aging Klotho Gene in Cancer Cell Lines. J Cancer Sci Ther 10: 351-357. doi:
10.4172/1948-5956.1000567
5. Zhou X, Wang X (2014) Klotho: A novel biomarker for cancer. J Cancer Res
Clin Oncol 141: 961-969.
6. Kurosu H, Kuro OM (2009) The klotho gene family as a regulator of endocrine
fibroblast growth factors. Mol Cell Endocrinol 299: 72-78.
7. Chen CD, Li H, Liang J, Hixson K, Zeldich E, et al. (2014) The anti-aging
and tumor suppressor protein klotho enhances differentiation of a human
oligodendrocytic hybrid cell line. J Mol Neurosci 55: 76-90.
8. Wolf I, Laitman Y, Rubinek T, Abramovitz L, Novikov I, et al. (2009) Functional
variant of klotho: A breast cancer risk modifier among BRCA1 mutation carriers
of Ashkenazi origin. Oncogene 29: 26-33.
9. Zhang H, Li Y, Fan Y, Wu J, Zhao B, et al. (2008) Klotho is a target gene of
PPAR-gamma. Kidney Int 74: 732-739.
10. Forster RE, Jurutka PW, Hsieh JC, Haussler CA, Lowmiller CL, et al. (2011)
Vitamin D receptor controls the expression of the anti-aging klotho gene in
mouse and human renal cells. Biochem Biophys Res Commun 414: 557-562.
11. Hsu SC, Huang SM, Lin SH, Ka SM, Chen A, et al. (2014) Testosterone
increases renal anti-aging klotho gene expression via the androgen receptormediated pathway. Biochem J 464: 221-229.
12. Hsu SC, Huang SM, Chen A, Sun CY, Lin SH, et al. (2014) Resveratrol
increases anti-aging Klotho gene expression via the activating transcription
factor 3/c- Jun complex-mediated signaling pathway. Int J Biochem Cell Biol
53: 361-371.
13. Sopjani M (2014) Relevance of the aging suppressor protein Klotho in health
and disease: Introduction, cellular signaling, mechanisms, clinical relevance,
conclusions, and perspectives. Lap Lambert Academic Publishing 2: 1.
23. Haussler MR, Whitfield GK, Haussler CA, Sabir MS, Khan Z, et al. (2016) 1,
25-Dihydroxyvitamin D and Klotho: A tale of two renal hormones coming of age.
Vitam Horm 100: 165-230.
24. Lau WL, Leaf EM, Hu MC, Takeno MM, Kuro OM, et al. (2012) Vitamin D
receptor agonists increase klotho and osteopontin while decreasing aortic
calcification in mice with chronic kidney disease fed a high phosphate diet.
Kidney Int 82: 1261-1270.
25. Kollitz EM, Zhang G, Hawkins MB, Whitfield GK, Reif DM, et al. (2016)
Evolutionary and functional diversification of the vitamin D receptor-Lithocholic
acid partnership. PLoS ONE 11: e0168278.
26. Forster RE, Jurutka PW, Hsieh JC, Haussler CA, Lowmiller CL, et al. (2011)
Vitamin D receptor controls expression of the anti-aging klotho gene in mouse
and human renal cells. Biochem Biophys Res Commun 414: 557-562.
27. Kolek OI, Hines ER, Jones MD, LeSueur LK, Lipko MA, et al. (2005) 1 alpha,
25-Dihydroxyvitamin D3 upregulates FGF23 gene expression in bone: The final
link in a renal-gastrointestinal-skeletal axis that controls phosphate transport.
Am J Physiol Gastrointest Liver Physiol 289: G1036-G1042.
28. Gattineni J, Twombley K, Goetz R, Mohammadi M, Baum M, et al. (2011)
Regulation of serum 1, 25 (OH)2 Vitamin D3 levels by fibroblast growth factor
23 is mediated by FGF receptors 3 and 4. Am J Physiol Renal Physiol 301:
F371-377.
29. Raghavan PR (2014) U.S Patent 8,722,093
30. Raghavan PR (2015) U.S Patent 9,006,292
31. Neubig RR (2007) Missing links: Mechanisms of protean agonism. Mol
Pharmacol 71: 200-1202.
14. Tang X, Wang Y, Fan Z, Ji G, Wang M, et al. (2016) Klotho: A tumor suppressor
and modulator of the Wnt/β-catenin pathway in human hepatocellular
carcinoma. Lab Invest 96: 197-205.
32. Panossian A, Wikman G (2009) Evidence-based efficacy of adaptogens in
fatigue, and molecular mechanisms related to their stress-protective activity.
Current Clin Pharmacol 4: 198-219.
15. Ligumsky H, Rubinek T, Merenbakh-Lamin K, Yeheskel A, Sertchook R, et
al. (2015) Tumor suppressor activity of klotho in breast cancer is revealed by
structure-function analysis. Mol Cancer Res 13: 1398-1407.
33. Raghavan PR (2017) Metadichol: A novel ROR gamma inverse agonist and its
applications in psoriasis. J Clin Exp Dermatol Res 8: 433.
16. Wang Y, Chen L, Huang G, He D, He J, et al. (2013) Klotho sensitizes human
lung cancer cell line to cisplatin via PI3k/Akt pathway. PLOS One 8: e57391.
17. Abramovitz L, Rubinek T, Ligumsky H, Bose S, Barshack I, et al. (2012) KL1
internal repeat mediates klotho tumor suppressor activities and inhibits bFGF
and IGF-1 signaling in pancreatic cancer. Clin Cancer Res 17: 4254-4266.
18. Xie B, Zhou J, Shu G, Liu DC, Zhou J, et al. (2013) Restoration of klotho gene
expression induces apoptosis and autophagy in gastric cancer cells: tumor
suppressive role of klotho in gastric cancer. Cancer Cell Int 13: 18.
19. Doi S, Zou Y, Togao O, Pastor JV, John GB, et al. (2011) Klotho inhibits
transforming growth factor-beta1 (TGF-beta1) signaling and suppresses renal
fibrosis and cancer metastasis in mice. J Biol Chem 286: 8655-8665.
20. King GD, Chen C, Huang MM, Zeldich E, Brazee PL, et al. (2012) Identification of
novel small molecules that elevate Klotho expression. Biochem J 441: 453-461.
34. Raghavan PR (2017) Metadichol® and vitamin C increase in vivo, an open-label
study. Vitam Miner 6: 163.
35. Raghavan PR (2017) Rheumatoid arthritis and osteoporosis: A case study. J
Arthritis 6: 240.
36. Raghavan PR (2017) Systolic and diastolic BP control in metabolic syndrome
patients with Metadichol® a novel nano emulsion lipid. J Cardiol & Cardiovasc
Ther 5: 555660.
37. Alemán CL, Más R, Hernández C, Rodeiro I, Cerejido E, et al. (1994) A
12-month study of policosanol oral toxicity in Sprague Dawley rats. Toxicol Lett
70: 77-87.
38. Alemán CL, Más Ferreiro R, Noa Puig M, Rodeiro Guerra I, Hernández Ortega
C, et al. (1994) Carcinogenicity of policosanol in Sprague-Dawley rats: A
24-month study. Teratog Carcinog Mutagen 14: 239-249.
21. Marco R (2017) A novel approach to klotho aimed at delaying and reversing
aging. BAOJ HIV 3: 029.
39. Alemán CL, Puig MN, Elías EC, Ortega CH, Guerra IR, et al. (1995)
Carcinogenicity of policosanol in mice: An 18-month study. Food Chem Toxicol
33: 573-578.
22. Xuan NT, Trang PT, Phong NV, Toan NL, Trung DM, et al. (2016) Klotho
sensitive regulation of dendritic cell functions by vitamin E. Biol Res 49: 45.
40. Georgiou A, Lisacek-Kiosoglous A, Yiallouris A, Stephanou A, Patrikios I (2017)
Klotho: The protein of faith. EC Neurology 75: 189-223.
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ISSN: 1948-5956
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