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Diluted Lycopodium Induced Cell Death and Clinical Improvement in

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Research Article
Diluted Lycopodium Induced Cell Death and Clinical Improvement in
Hepatocellular Carcinoma
Debasmita Chatterjee 1, Banhishikha Singh 1, Ashoke Kumar Pradhan 2, Krishnendu Paira1 and Satadal Das1*
1
Tissue Culture Unit, Department of Biotechnology, Heritage Institute of Technology, Kolkata, West Bengal, India
2
Institute of HYDT Research & Education, Kolkata, West Bengal, India

ARTICLE INF O ABSTRACT

Article history: Hepatocellular carcinoma (HCC) is a common cancer with high incidence rate, and 5-year survival rate in
Received: 15 June, 2022 HCC is less than 20%. Thus, in search of newer anticancer agents effective in HCC, we have explored
Accepted: 27 June, 2022 possible usefulness of an alternative medicine Lycopodium against the human liver cancer cell line, HepG2
Published: 8 July, 2022 along with its clinical efficacy. The HepG2 cell line was challenged with Lycopodium 6C (diluted
Keywords: Lycopodium <1pg /mL available as alternative medicine) along with vehicle alcohol control in 24 hours.
Hepatocellular carcinoma The cytopathic effect and viability test with methylene blue stain were observed. The cells were harvested
Lycopodium for total RNA extraction, and gene expression levels of targeted cytokines -Interferon gamma (IFN γ);
cytokines Interleukins - IL-6, IL-8, IL-10, IL-1β, Transforming Growth Factor- TGF-β1, TGF-β3 and Tumor Necrosis
gene expression Factor alpha (TNF-α) by RT-PCR were studied. DNA fragmentation assay and cell viability assay by MTT
anti-cancer agent method were also tested. After ethical permission we applied this medicine as adjunct therapy to observe
any beneficial role of the medicine. Statistically significant changes of IL-10, IL-1β and TGF-β3 were
observed after challenge with Lycopodium 6C. The IL-10 gene expression in malignant cells was
significantly reduced with Lycopodium 6C; however, the expression is more with vehicle alcohol compared
to normal control set. Thus, the medicine could decrease the excessive IL-10 gene expression to a moderate
level. IL-1β and TGF-β3 gene up-regulation by the vehicle alcohol were also mitigated by the medicine
Lycopodium 6C. Mild DNA fragmentation was also seen in cancer cells after challenge with the medicine.
Two cases suffering from hepatocellular carcinoma showed much clinical improvement after therapy with
this medicine. Lycopodium 6C may act as a supporting alternative medication for treating HCC.

© 2022 Satadal Das. Hosting by Science Repository.

Among the other modes of treatment, homeopathy is considered to be an

Introduction
alternative treatment approach with no such significant side effects and
also lessens the pain associated with the disease [4]. The homeopathic
Cancer is a deadly disease that has already taken the entire world in its
clinicians prescribes both mother tinctures and potentized preparations
grip, and so the problem requires a multifaceted approach for its
for the treatment purpose depending on the symptomatic manifestations
treatment [1]. Among the varied forms of cancer, hepatocellular
of the patient and have claimed that both mother tincture and potentized
carcinoma (HCC) is very common due to its high frequency and
preparations are useful against the disease condition [4]. Researchers
incidence rate. A chronic inflammatory environment induces the
have also revealed that the homeopathic medicine Lycopodium could
development of HCC which usually represent the chronic active hepatitis
reduce the pathogenesis associated with the tumor like growth, and in
following viral infection or due to substance abuse (alcohol) [1, 2].
turn could also retard the growth of the tumor itself. Moreover, the
Surgical resection and/or transplantation are considered to be the two
homeopathic preparation was evident to enhance the life span of the
most efficient treatment methodologies for liver cancer, but
tumor carrying animals [4, 5].
unfortunately not all patients could avail these treatment procedures [3].

*Correspondence to: Satadal Das, Tissue Culture Unit, Department of Biotechnology, Heritage Institute of Technology, Kolkata, West Bengal, India; ORCID: 0000-
0002-9843-3466; E-mail: [email protected], [email protected]

© 2022 Satadal Das. 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. Hosting by Science Repository.
http://dx.doi.org/10.31487/j.COR.2022.03.02
Diluted Lycopodium Induced Cell Death and Clinical Improvement in Hepatocellular Carcinoma 2

In our study we have chosen Lycopodium 6C against the HCC cell line medium was decanted and fresh media was added of required volume to
HepG2. Lycopodium is also known as “clubmosses”, “creeping cedars” carry out the experiment in a 6wells plate [6, 7].
and “ground pines” and the plant belongs to family “Lycopodiaceae” [6].
The medicine showed varied uses in homeopathy such as for 1 mL of cells containing media was added to each well of the plate and
constipation, chronic lung and bronchial disorders, fever, aneurysms and kept in 5% carbon dioxide incubator in a humidified environment for the
also against cancer [6]. It can also be useful against gastric inflammation next 24 hours. The next day the wells were washed with 1X PBS and
and chronic kidney disease. Several researchers have also claimed about again fresh media was added. The cells were allowed to reach a
its hepato-protective, anti-oxidant, immune modulatory, confluency of 10 5 - 106 before the inoculation of the inoculums [6, 7].
neuroprotective, antimicrobial, analgesic, and anti-cancer properties [6].
Thus in the present study we have explored the anti-cancer activity of IV Inoculation of Medicine
the homeopathic preparation “Lycopodium 6C” against the human liver
cancer cell line (HepG2). The cells were challenged with 100 µL of medicine, Lycopodium 6C to
three wells each (triplicate sets), Alcohol vehicle in another three wells
Materials And Methods and the rest three wells served as control without any inoculums. The
plates were rotated clockwise and anti-clockwise for mixing of the
I Procurement of Cell line inoculums. Immediately after inoculation pictures of cell line were taken
(0 hour after inoculation).
The HepG2 was procured from National Centre for Cell Science, Pune,
India. The cells was transported in a T25 cm 2 flask within growth V Modified Methylene Blue Assay
medium, Minimum essential medium (Eagle) with 2mM L-glutamine
and Earle’s Balanced Salt Solution (BSS) that was adjusted to contain Modified methylene blue assay can be used for a wide range of cell
1.5g/L sodium bicarbonate, 0.1mM non-essential amino acids and counting process. This method is advantageous compared to other
1.0mM sodium pyruvate, 90% concentrated fetal bovine serum of final staining method because the stain can be applied directly upon the
concentration of 10%. adherent cell lines that are growing in any size culture plates [8]. The
methylene blue solution is prepared with Phosphate buffer saline
II Procurement of Chemicals solution (1X, PBS), 1.25% glutaraldehyde, and 0.06% methylene blue.
The stain was added of volume 1 ml to each well of the 6 well culture
The medicine Lycopodium 6C was purchased from “HAPCO, India” - plate, and it was incubated at 37ºC for one hour. Then the methylene blue
an India government recognized homeopathic medicine producing solution was thoroughly rinsed with PBS solution and then pictures were
company”. It was a diluted and potentized alcoholic plant extract taken under inverted microscope to differentiate among the live and dead
material prepared by homeopathic pharmacopoeia guidelines. The cells. The cells were observed under inverted microscope under 20X,
Dulbecco’s Modified Eagle Medium (DMEM) (1X) along with and 40X to record the cytopathic effect [8].
Glutamax was procured from Gibco, ThermoFischer, USA. The medium
was supplemented with F-12 (1X) nutrient mixture Ham + L-glutamine VI Gene Expression Analysis of Cytokines
(Gibco, ThermoFischer, USA) for the better growth of the HepG2 cell
line. The Fetal bovine serum (FBS) and the antibiotic –antimycotic The cells of each well were harvested with 1ml RNA isoplus and the
solution namely Penicillin/Streptomycin/Amphotericin B Solution total RNA was extracted following the instructions of the manufacturer.
(100X), Phosphate buffer saline (PBS, 1X) of pH 7.4, Trypsin enzyme The total RNA was estimated using A260/280 ratio and the cDNA were
(0.05 X) were all purchased from Gibco, Thermo Fischer, USA. The synthesized using the reverse transcriptase kit (Bio-Rad, USA). Then
MTT assay kit EZ Count was purchased from Himedia, India. The 2µL of the cDNA was utilized for SyBR green RT PCR assay (Bio-Rad,
molecular biology chemicals such as the RNA isoplus were purchased USA) in CFX-96 model of RT PCR, Bio-Rad, USA. The gene
from Takara, the cDNA synthesis reverse transcriptase kit and the iTaq expression analysis was conducted of the following genes namely,
Sybr green supermixture for RT-PCR were purchased from Bio-Rad, Interferon (IFN) gamma, Interleukins- IL-6, IL-8, IL-10, IL-1β,
USA. Transforming growth factors (TGF) – β1 and β3, and Tumor necrosis
factor alpha (TNF α) against the housekeeping gene, β actin [9, 10].
III Cell Culture
VII MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium
The HepG2 cells were allowed to reach 80% confluency in a T25 cm 2 bromide) Assay for Cell Viability
flask for 48 hours with DMEM+F12 supplemented with antibiotic
solution, and 10% FBS media for 48 – 72 hours [7]. The media change The HepG2 cells viability was measured using the MTT assay kit EZ
was given at a regular interval of 48 hours after washing the cellular Count (Himedia Pvt. Ltd., India). The assay was done keeping the
debris with PBS (1X). The cells were splitted using Trypsin solution for required controls such as medium control (without HepG2 cells), Cell
10 minutes and then the flask was agitated manually, so that the adherent line control (Media + HepG2 Cells), and the vehicle control (Medium
cells leave the base of the flask. Immediately after 10 minutes, the trypsin containing the vehicle solvent or the experimental drug). The volume of
was inactivated with DMEM media supplemented with FBS (10%). The experimental drug inoculated upon the cell line was 100µL and the
cells were then centrifuged at 1200 rpm for 12 minutes. The supernatant culture plate was incubated at 37ºC for one hour in carbon dioxide and
humidified atmosphere. Following incubation, 10µL of MTT reagent

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Diluted Lycopodium Induced Cell Death and Clinical Improvement in Hepatocellular Carcinoma 3

was added to the experimental sets and control sets. Thereafter, the plate phenol-choloroform extraction. After extraction the isolated DNA was
was incubated in the above-mentioned condition for the next 4 hours. dissolved in 60µL of elution buffer and the purity of the DNA was
The plate was observed at regular intervals under the inverted checked at A260/A280 ratio using UV-Vis Spectrophotometer (Agilent,
microscope as it would develop formazan crystals. After the formation Singapore). Absorbance of 1.7 was considered to be of pure isolated
of crystals, 100µL of the solubilization solution was added to all the DNA.
wells, and the plate was shaken by hand for about 5 minutes, and then
the plate was kept within the incubator for overnight incubation. The Then the whole genomic DNA of all the samples were loaded within 1%
crystals would dissolve within the overnight incubation, and the agarose gel for comparison of the intact DNA band of control with the
coloration would develop. Then the absorbance was taken at 570 nm by medicine and alcohol control counterparts. Then the gel was stained
an automated 96 well plate ELISA reader (Roboniks, India). The data using ethidium bromide for band visualization and photography using
was graphically recorded [11]. UV-Vis trans-illuminator [12].

VIII DNA Fragmentation Assay IX Application in Patients Suffering from Hepatocellular

Carcinoma
The phenomenon of DNA fragmentation and condensation of nucleus
contents are considered to be the activities to be taking place during late After getting ethical permission we could apply this medicine in two
apoptosis. Endonuclease which is located at the margins of nucleus can cases suffering from hepatocellular carcinoma as adjunct therapy with
break the chromatin DNA into short fragments during the apoptosis. consent taken from the patients.
Here in our experiment, the cells of each well were trypsinized and cell
scrapper was used to collect maximum number of cells from each wells. X Statistical Analysis
Then the cells were washed with PBS twice and proteinase K was added
to each tube and incubated at 56ºC for 1 hour at water-bath [12]. Then The data was analysed for statistical significance using one-way analysis
the whole genomic DNA was isolated following the standard protocol of of variance (ANOVA) using statistical software, GraphPad Prism 9.3.1.

Figure 1: Cell cytopatheic effect before and after methylene blue assay - In the control set the HepG2 cells were in usual shape and size along with intact
outline. The inoculums were added when confluency was around 60% to 80%. In the medicine set, the cells were round in shape indicating apoptosis.
Maximum cells were dead and were uplifted from the base (detachment of cells) of the culture plate after 24 hours of the inoculation. In case of alcohol
(vehicle control set), the cells also showed apoptosis, i.e., rounding of cells and cell detachment from base of the culture plate. The size of the cells decreased
considerably; however, some cells were still alive in decreased size and were attached to the base of the culture plates. A) Represents control HepG2 cell
line (after 24 hours). B) Cell line after 24 hours of drug inoculation, Lycopodium 6C. C) Cell line after 24 hours of inoculation of vehicle control (alcohol).
D) Control HepG2 cells after 24 hours after methylene blue staining. E) Cell line after 24 hours of drug inoculation, Lycopodium 6C after methylene blue
staining. F) cells after 24 hours of inoculation with vehicle control (alcohol) after methylene blue staining.

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Diluted Lycopodium Induced Cell Death and Clinical Improvement in Hepatocellular Carcinoma 4

Results considerably; however, some cells were still alive in decreased size, and
were attached to the base of the culture plates. After methylene blue
I Cell Morphology and Cytopathic Effect staining, the control cells were stained with intact cellular morphology,
usual size and intact margins, whereas in case of alcohol (vehicle
In the control set the HepG2 cells were in usual shape and size along control) the cells were decreased in size with membrane blebs. However,
with intact outline. The inoculums were added when cells were around few cells had retained the stain indicating that the cells were still alive.
80% confluent. In the medicine set, the cells became small round in In the medicine set, the cells were all rounded in shape and were floating
shape indicating possibility of apoptosis. Maximum cells were dead and within the media, indicating that all the cells have undergone apoptosis
were uplifted from the base (detachment of cells) of the culture plate or necrosis (Figures 1A-1F).
after 24 hours of the inoculation. In case of alcohol (vehicle control set),
the cells also showed apoptosis, i.e., rounding of cells and cell
detachment from base of the culture plate. The size of the cells decreased

Figure 2: Differential expressions of IFN γ and IL-6 genes in different experimental sets – the bar graphs represents the mean value ± SEM of three
independent experimental findings A) IFN γ was found decreased in the medicine set in comparison to the alcohol and normal control. B) In case of cytokine
IL -6, the expression of this particular gene was found to be mildly high in the alcohol and normal control, but on the contrary the gene expression of IL-6
was decreased by the Lycopodium 6C set. The change is not statistically significant.

Figure 3: Differential expressions of IL-10 and IL-8 genes in different experimental sets – the bar graphs represents the mean value ± SEM of three
independent experimental findings A) the observation was noted for the IL -10 where the medicine diminished the gene expression in that set, however, the
gene expression of this particular cytokine is high within the alcohol control with respect to the normal control set. B) However, in case of the pro-
inflammatory cytokine, IL -8 the values were enhanced by the medicine, Lycopodium 6C set and alcohol control in comparison to its normal control.

II Differential Expressions of Cytokine Genes Using RT PCR the gene expression of IL-6 was decreased by the Lycopodium 6C set.
Technique The same observation was noted for the IL -10 where the medicine
diminished the gene expression in that set, however, the gene expression
The differential gene expressions of the selected cytokines were of this particular cytokine was high within the alcohol control, with
represented in the bar diagrams (Figures 2-5). IFN γ was found decreased respect to the normal control set. However, in case of the pro-
in the medicine set in comparison to the alcohol and normal control. In inflammatory cytokine, IL -8 the values were enhanced by the medicine,
case of cytokine IL -6, the expression of this particular gene was found Lycopodium 6C set and alcohol control in comparison to its normal
to be mildly high in the alcohol and normal control, but on the contrary control. IL-1 β which is considered to be a key mediator in the

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Diluted Lycopodium Induced Cell Death and Clinical Improvement in Hepatocellular Carcinoma 5

inflammation process was found to be high in the alcohol control set normal control set, the value was 1.20. The last parameter was Tumor
when compared to normal control, and the value was much reduced by necrosis factor α (TNF α), was found to be increased in the alcohol set
the medicine in that set. TGF, which exhibits anti-tumerogenic effect in and reduced in the medicine set when compared to control. MTT assay
the initial stage by induction of cytostatis and apoptosis was found to be revealed that more number of HepG2 cells were alive when compared to
decreased in the alcohol control and medicine set with respect to control. alcohol vehicle control at the selected drug doses of 10 to 120 µL (Figure
However, TGF β3 gene expression showed marked increase in the 6). The DNA fragmentation assay also revealed mild fragmentation of
alcohol control and it was reduced in the medicine set whereas in the DNA (Figure 7).

Figure 4: Differential expressions of IL-1β and TGF-β1 genes in different experimental sets – the bar graphs represents the mean value ± SEM of three
independent experimental findings. A) IL-1 β which is considered to be a key mediator in the inflammation process was found to be high in the alcohol
control set when compared to normal control and the value was much reduced by the medicine in that set. B) TGF, which exhibits anti-tumerogenic effect
in the initial stage by induction of cytostatis and apoptosis was found to be decreased in the alcohol control and medicine set with respect to control.

Figure 5: Differential expressions of TNF-α and TGF-β3 genes in different experimental sets – the bar graphs represents the mean value ± SEM of three
independent experimental findings A) Tumor necrosis factor α (TNF α), was found to be increased in the alcohol set and reduced in the medicine set when
compared to control. B) However, TGF β3 gene expression showed marked increase in the alcohol control and it was reduced in the medicine set whereas
in the normal control set, the value was 1.20.

Figure 6: MTT assay curve of Lycopodium 6C and alcohol - MTT assay revealed that more number of HepG2 cells were alive when compared to alcohol
vehicle control at the selected drug doses of 10 to 120 µL.

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Diluted Lycopodium Induced Cell Death and Clinical Improvement in Hepatocellular Carcinoma 6

Figure 7: DNA Fragmentation Assay -The DNA fragmentation assay also revealed mild fragmentation of DNA.

III Clinical Study Results IV Statistical Analysis

Both the cases showed significant clinical improvement after treatment The data showed statistical significance for the cytokine parameters, IL-
with the medicine. Details of those two patients are given in the (Table 10, IL-1β and TGF-β3 with P-value less than equal to 0.05. The other set
1). Both the cases were males and they were hepatitis B positive with values are mentioned in the (Table 2).
markedly increased AFP. Pain, nausea, vomiting, anorexia, tenderness
of abdomen, and weakness relieved 1-2 months after administration of
the medicine.

Table 1: Different parameters of the two cases those were treated with Lycopodium.
Report / Findings
SGPT (U/mL)

CEA (ng/mL)

AFP (ng/mL)

Remark (s)
Hb (G/dL)

Bilirubin
(mg/mL)
Age (Yr)

CA 19.9

CA 72.4
(U/mL)

(U/mL)
HBsAg
FNAC
Name

USG
Sex

NB 37 M HCC Ascites, Mild 13.8 0.7 62 + <0.5 13895.4 24.31 0.31 Clinical improvement
hepato- for 2 months after
splenomegaly, giving two doses of
mesenteric the medicine. Pain,
lymphadenitis nausea, vomiting,
anorexia relieved.
HGR 68 M HCC Hepato- 11.7 1.06 28 + 3.4 340 68.04 0.77 Clinical improvement
splenomegaly, for 1 month after
SOL in right lobe, giving one dose of the
portal vein medicine. Pain,
thrombosis, nausea, vomiting,
periportal tenderness of
lymphadenopathy abdomen, severe
weakness relieved.

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Diluted Lycopodium Induced Cell Death and Clinical Improvement in Hepatocellular Carcinoma 7

Table 2: Statistical Analysis ANOVA summary table of the gene expression analysis values.
SL. No. Set Analysed F value P-value R squared value
(A-C)
1 IFN γ 2.769 2.083 (NS) 0.6486
2 IL-6 1.666 0.3261 (NS) 0.5263
3 IL-8 0.8445 0.5118 (NS) 0.3602
4 IL-10 17.52 0.0221 (S)* 0.9212
5 IL-1β 72.61 0.0029 (S)* 0.9798
6 TGF-β1 2.425 0.2363 (NS) 0.6178
7 TGF-β3 73.41 0.0028 (S)* 0.9798
8 TNF-α 2.674 0.2155 (NS) 0.6406
*Significant at 0.05 level.
s: significant; ns: non-significant.

Discussion The cytokine IL-8 was also found to be high among liver cancer tissues
and it was evident clinically that there is metastasis with elevated
It is well documented that IFN γ plays a significant role in the host frequency of portal vein, venous and bile duct invasions [18]. The
defense procedure but it is still unclear that in what mechanisms does researchers also confirmed that IL-8 serves as an angiogenesis factor in
HCC evades or blocks the signal transduction mechanism of immune case of HCC and plays a significant role in the metastasis and invasion
supervision of IFN- γ [13]. It was observed within a clinical data that the of HCC [18]. However, in experimental findings we found there is up-
expression of IFN- γ receptors on the surface of cells was induced or regulation of expression of IL-8 gene in the medicine set when compared
stimulated among 27 non –cancerous liver tissue samples. On the with respect to alcohol and control sets. Several authors have conferred
contrary, in case of non-stimulated IFN- γ receptors, the size of the tumor the activities of pro and anti-tumerogenic activity to cytokine IL-1 and
was large (statistically significant, P = 0.032) along with higher serum its family [19]. We could see an up-regulation of IL-1β in the alcohol set
alpha-fetoprotein (AFP) level (P value = 0.001) [13]. However, in but the expression level has been controlled by the medicine Lycopodium
another research study, IFN- γ has a significant role in the induction of 6C. Transforming growth factor beta is evident to play role to inhibit the
anti-tumerogenic response [14]. The anti-tumerogenic activity of IFN- γ growth of tumor in the early stage of liver cancer with the induction of
is based on the following functions – anti-tumerous, pro-apoptotic and cytostasis and apoptosis, however, can promote malignant cases in their
cytostatic activities and due to these activities, it has a significant role in advanced stages [20]. The gene expression study revealed that the value
adjuvant immune-therapy against varied forms of cancers. However, the of TGF beta 3 is high in the alcohol set which got ameliorated in the
researchers have confirmed that the resulting concentration of IFN- γ in medicine set. The last and the most important factor is tumor necrosis
the micro-environment of the tumor determines its anticancer role. factor alpha (TNF α) which is evident from research study to promote
the growth of tumor along with poor prognosis of HCC [21]. Our study
In our data, we could observe that gene expression of IFN- γ has revealed that alcohol increased the level of gene expression of TNF α
decreased up to 0.02 fold with respect to the housekeeping gene, β-actin which got reduced with the application of medicine, Lycopodium 6C
[14]. The role of IL-6 is multifunctional that demonstrates a varied types when compared to control.
of activities in different pathological conditions [15]. The main activities
of this cytokine can be studied in the environment of liver, where it is Thus, all the cytokine expression changes studied in this experiment
mainly produced. Group researchers studied its clinical prognostic role appears beneficial to the patient with liver cancer except IL-8 gene
in case of HCC and found a significant positive correlation of IL-6 serum expression. However, raised 10.48-fold cytokine gene expression in
concentration and tumor size among the HCC subjects. The study vitro is not so significant, and it is usually counteracted by other
concluded that IL-6 could help in progression of HCC by acting as an cytokines which may lead to a negative action on the cancer cells leading
autocrine tumoral growth factor and in turn reducing immune to a possible remission of cancer in vivo as a whole. Our preliminary
supervision [15]. Our data indicated that Lycopodium 6C could control observation with two cases of hepatocellular carcinoma showed
the gene expression of IL-6 thus limiting its role in tumor progression. symptomatic improvements after administration of this medicine.
Another group of researcher also studied the mean serum level of
cytokine IL-10 among the HCC subjects via ELISA method and Acknowledgement
concluded that the level of IL-10 is quite high and it can act as a
biomarker along with AFP and IL-6 for the patient [16]. It was also The authors would like to acknowledge Dr Sajjan Bhajanka, Trustee
mentioned by other researchers that high level of IL-10 correlates with member, Kalyan Bharati Trust and Shri Pradip Agarwal, Chief
worse prognosis of patients with negative survival rate suffering from Executive Officer of Heritage Institute of Technology to grant
varied forms of cancer [17]. In our data, the value of IL-10 was found to permission and provide infrastructure for conducting this research study.
be reduced in the Lycopodium 6C set, however, the value was higher in
the alcohol set when compared to normal control set. Thus, the medicine Funding
could control the high gene expression level of IL-10 which is directly
correlated with worsening of the pathophysiological condition. None.

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 Diluted Lycopodium Induced Cell Death and Clinical Improvement in Hepatocellular Carcinoma 8

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