Total saponins in Rubus parvifolius L. induce lymphoma cells apoptosis through upregulated Bax/Fas and downregulated Bcl-2 in vivo and in vitro

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Submit a manuscript: https://www.tmrjournals.com/tmr TMR | March 2019 | vol. 4 | no. 2 |
doi: 10.12032/TMR20190304108
Traditional Chinese Medicine
Total saponins in Rubus parvifolius L. induce lymphoma cells
apoptosis through upregulated Bax/Fas and downregulated Bcl-2 in
vivo and in vitro
Xiao-Feng Xu1
*, Ru-Bin Cheng2
, Xue-Jin Zhang1
, Rui-Lan Gao3
1
Department of Hematology, Zhejiang Provincial Integrated Chinese and Western medicine Hospital, Hangzhou,
China. 2
College of Pharmaceutical Science, Zhejiang Chinese Medical University, Hangzhou, China. 3
The First
Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou, China.
*Corresponding to: Xiao-Feng Xu, Department of Hematology, Zhejiang Provincial Integrated Chinese and
Western medicine Hospital, 208, Huancheng East Road, Xiacheng District, Hangzhou, China. Email:
hhxuxiaofeng@126.com.
Highlights
Total saponins of Rubus parvifolius L. (TSRP) induces Raji cell apoptosis by inhibiting Bcl-2, increasing
Bax expression, and reducing Fas expression in vivo and in vitro. Additionally, TSRP has some effects on
the chondriosome pathway of apoptosis in vitro.
Traditionality
Rubus parvifolius L. (RP) belongs to the family Rosaceae and is mainly produced in the temperate zone of
the Northern hemisphere. As early as the Xihan Dynasty of China, Erya recorded that RP were edible.
Bencao Shiyi recorded the medicinal properties of the root bark and fruit of RP, which include clearing heat,
cooling blood, stopping bleeding, dispersing knots, relieving pain, diuresis, and detumescence.

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doi: 10.12032/TMR20190304108
Abstract
Purpose: To investigate the effect of total saponins of Rubus parvifolius L. (TSRP) on lymphoma Raji cells and
further discuss its mechanism. Methods: The model of nude mice bearing Raji cells was established, the volume,
weight and inhibition rate of the transplanted tumor were analyzed and compared after different concentrations of
TSRP treatment. Cell apoptosis and expression of Bcl-2, Bax, Fas proteins were detected by TUNEL and
immunohistochemiscal method respectively. Effects of TSRP on cell proliferation were tested with MTT assay in
vitro. Cell apoptosis and expression of Caspase-9, Caspase-3, Bcl-2, Bax and Fas proteins were tested with DAPI
staining and Western blot. Results: TSRP significantly reduced the volume and tumor weight of Raji subcutaneous
transplanted tumor and induced the apoptosis of Raji cells in vivo. The tumor inhibition rate of high-dose (100
mg/kg) TSRP is 90.84%. The TUNEL test results show that the fluorescence intensity of the tumor issue treated
with TSRP is significantly improved. Compared with the control group, the fluorescence intensity of
high-concentration TSRP is 82.43 ± 7.81, which is significantly different (P < 0.001). The results of
immunohistochemistry test showed that the Bcl-2 expression of Raji cell treated with TSRP is obviously reduced,
and Bax expression is obviously increased. Meanwhile, compared with that of control group, Fas expression is
obviously reduced. MTT assay showed that TSRP can significantly inhibit proliferation of Raji cells with dose
dependence. The inhibition rate of 400 μg/mL TSRP is 53.46 ± 4.90% (P < 0.001). DAPI staining results showed
that TSRP can significantly induce cell apoptosis. According to Western blot results, it is found that TSRP can
significantly inhibit activity of Bcl-2 and increase Bax expression, and TSRP can also inhibit Fas expression.
Meanwhile, expression of Caspase-9 and Caspase-3 is also increased. Conclusion: TSRP could inhibit the
proliferation of lymphoma via induction of apoptosis in a time and dose-dependent manner. Apoptotic signaling
induced by TSRP was characterized by up-regulating Bax, Fas and Caspase-8 protein expression, and
down-regulating of Bcl-2 protein expression.
Keywords: Total Saponins of Rubus parvifolius L., Bcl-2, Bax, Fas, Apoptosis, Lymphoma
Abbreviations:
RP, Rubus parvifolius L.; TSRP, Total saponins of Rubus parvifolius L.; TCM, Traditional Chinese medicine;
IMDM, Iscove’s Modified Dulbecco’s Medium; STAT-3, Signal transducer and activator of transcription 3;
JNK, c-Jun N-terminal kinase; AMPK, Monophosphate- activated protein kinase; Akt, Protein kinase B;
mTOR, Mammalian target of rapamycin; CAR-T, Chimeric antigen receptor T-Cell immunotherapy; DAB, 3,
3-Diaminobenzidine; TBST, Tris buffered saline Tween.
Acknowledgments:
The study were supported by grants from Zhejiang Provincial Administration of Traditional Chinese Medicine
(No. 2011ZA081, 2013ZB095 and 2015ZA147), Hangzhou Medical Science and Technology Plan (No.
2012A048).
Competing interests:
The authors declare that there is no conflict of interests regarding the publication of this paper.
Citation:
Xiao-Feng Xu, Ru-Bin Cheng, Xue-Jin Zhang, et al. Total saponins in Rubus parvifolius L. induce lymphoma
cells apoptosis through upregulated Bax/Fas and downregulated Bcl-2 in vivo and in vitro. Traditional
Medicine Research 2019, 4(2): 99-108.
Executive Editor: Cui-Hong Zhu.
Submitted: 28 December 2018, Accepted: 1 March 2019, Online: 4 March 2019.

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doi: 10.12032/TMR20190304108
Background
Rubus parvifolius L. (RP) belongs to the family
Rosaceae and is mainly produced in the temperate
zone of the Northern hemisphere. As early as the
Xihan Dynasty of China, Erya recorded that RP were
edible. Bencao Shiyi recorded the medicinal properties
of the root bark and fruit of RP, which include clearing
heat, cooling blood, stopping bleeding, dispersing
knots, relieving pain, diuresis, and detumescence.
Modern pharmacological experiments have shown that
the water extract of RP has hemostatic,
blood-activating, and stasis-removing effects and RP
has been successfully used to treat coronary heart
disease, angina pectoris, and other cardiovascular
diseases [1].
Our previous studies demonstrated that total
saponins of R. parvifolius L. (TSRP) can significantly
induce apoptosis in human chronic myeloid leukemia
K562 cells in vivo and in vitro. The mechanism
involves increasing the expression of poly ADP ribose
polymerase, caspase-3, and caspase-9 and inhibiting
Bcl-2 expression. Additionally, we showed that TSRP
can obviously inhibit the phosphorylation of Signal
transducer and activator of transcription 3 (STAT3) and
increase the phosphorylation of adenosine
monophosphate- activated protein kinase (AMPK) and
c-Jun N-terminal kinase (JNK). A high concentration
TSRP can significantly inhibit phosphorylation of
protein kinase B (Akt) and mammalian target of
rapamycin (mTOR) [2].
Lymphoma is a common tumor of the circulatory
system. Although great progress has been made in
treating lymphoma by the development of molecular
targeted therapy and chimeric antigen receptor T-Cell
immunotherapy (CAR-T), combined chemotherapy
remains the main treatment method. However, the
toxicity and drug resistance of chemotherapy drugs
remain as limitations. Few studies have examined the
inhibitory effect of TSRP on lymphoma. Zheng
Zhenxiao et al [3] reported that TSRP had anti-tumor
activity against Hut-78 human skin T cell lymphoma.
Through clinical observation, we found that oral
administration of RP can reduce fever and night sweats
in patients with lymphoma. Therefore, we examined
the effect of TSRP on lymphoma to provide a basis for
identifying the effective constituents in this traditional
Chinese medicine (TCM) that inhibit the malignant
proliferation of lymphoma cells.
Methods
Materials
Iscove’s modified Dulbecco’s medium (IMDM)
was purchased from GIBCO (Grand Island, NY, USA).
Antibodies against Bcl-2, Bax, Caspase-9, Caspase-3,
Fas and β-Actin were obtained from Cell Signaling
Technology (Danvers, MA, USA). All reagents were
used according to the manufacturer's instructions.
Cell culture
Raji cells were ordinarily inoculated in IMDM
containing 10% inactivated calf serum and 100 U/mL
each of penicillin and streptomycin. The culture
solutions were incubated in a constant temperature
tank (37℃, 5% CO2 and with saturated humidity) for
suspension culture. The medium was replaced every
2-3 days. Cells in the logarithmic growth phase were
collected for analysis.
TSRP
TSRP was provided by the TCM Resource
Engineering Lab of Zhejiang Chinese Medical
University. An acetic anhydride-concentrated sulfuric
acid was reacted with the available samples. The
results were positive, demonstrating that the obtained
substance was a triterpenoid saponin and that the
purity was above 90%. TSRP was dissolved in water at
a concentration of 1.0 g/L, sterilized by filtration, and
stored at 4℃ until use.
Establishment of nude mice model bearing
lymphoma and grouping
Raji lymphoma cells were centrifuged for 10 min at
1000 r/min, after which the supernatant was removed.
IMDM was used to prepare uniform mixtures of Raji
lymphoma cells. The cell concentration was 2 ×
107
/mL, and the Raji lymphoma cells were
subcutaneously transplanted into the left armpit of
3-week-old BALB/c nude mice. The dose was 0.2
mL/mouse; 20 mice were used in total. Tumors formed
after culture for 2 weeks (at that time, tumor nodules
were observed below the skin of all mice; the nodule
size was approximately 2 × 2 mm). Five nude mice did
not develop tumors, giving a tumor formation rate of
75%. The nude mice in which tumors had formed were
randomly divided into three groups, with each group
consisting of 5 mice: ① Control group (normal saline);
② TSRP low-dose group (20 mg/kg); and ③ TSRP
high-dose group (100 mg/kg).
Analysis of tumor inhibition by TSRP in nude mice
bearing lymphoma
For the TSRP low-dose and high-dose groups, 100 µL
was intragastrically administrated for each mouse
every day at concentrations of 4 and 20 mg/mL,
respectively. For the control group, the same volume of
normal saline was intragastrically administered to each
mouse every day. The activities (including eating,
properties of stool and urine, mental status, etc.) of the
mice were observed every day. Tumor size was
measured every three day. The longer diameter a and
shorter diameter b (mm) were measured with a Vernier
caliper and tumor volume was calculated as follows: V

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= π/6 × ab2
(mm3
) [4], after which the tumor growth
curve was drawn. After intragastric administration of
TSRP or saline for 3 weeks, the mice were sacrificed
by cervical dislocation. The tumor was removed and
weighed, and the tumor inhibition rate was calculated
as follows: tumor inhibition rate = (1 － tumor weight
in the treatment group/tumor weight in the control
group) × %.
TUNEL analysis
Paraffin sections containing the tumor tissue were
prepared. After antibody labeling of the tissue using
the TUNEL method, 50-100 μL freshly prepared 3,
3-Diaminobenzidine (DAB) solution as the
chromogenic substrate was added to each section, and
color changes were observed under a microscope.
After color development, the section was rinsed with
distilled water and restained with hematoxylin. Neutral
gel was used to mount the section and the section was
observed under a fluorescence microscope.
Immunohistochemistry analysis
Dewaxing, antigen retrieval, primary antibody
incubation, and secondary antibody incubation
(horseradish peroxidase-labeled) were conducted for
the sections. DAB solution was added as the
chromogenic substrate. If the result is positive, the
color was brown yellow. After restaining the sections
with hematoxylin, neutral gum was used to mount the
sections.
MTT assay
Lymphoma cells were cultured under normal oxygen
and low oxygen conditions. Single cell suspensions
were prepared with the culture solution containing
10% fetal calf serum. Next, 103
-104
cells were
inoculated into each well of a 96-well plate; the
volume of each well was 200 µL. Raji cells were
treated with 0, 100, 200, or 400 µg/mL TSRP. After
culturing the cells for 48 h, 20 µL MTT solution was
added to each well (5 mg/mL, prepared with PBS, pH
7.4) and incubation was continued for 4 h. The
supernatant in the wells was carefully suctioned and
discarded, after which 150 µL dimethyl sulfoxide was
added to each well. The samples were incubated with
shaking for 10 min to dissolve the crystals. Absorption
at a wavelength of 490 nm was measured in each well
with an enzyme-linked immune assay reader.
DAPI staining
Based on the results of our previous experiment [2], 0,
100, 200, and 400 µg/mL TSRP were used. Raji cells
in the logarithmic growth phase were cultured. The
cells were treated with TSRP for 24 h, rinsed with cold
PBS twice, and were fixed for 30 min with ethanol at
-20℃. The fixed Raji cells were rinsed with PBS,
stained for 15 min with 10 μg/mL DAPI at 25℃, and
then rinsed with PBS three times. Raji cells were
observed under a fluorescence microscope, and cells
containing concentrated chromatin or cracked nuclei
were considered as apoptotic.
Western blot analysis
Raji cells were treated with 0, 100, 200, and 400
µg/mL TSRP individually. Total protein was extracted,
and the sample concentration was measured. For
sodium dodecyl sulfate-polyacrylamide gel
electrophoresis, 20 µg sample was added to each well
of the gel. After electrophoresis, the proteins were
transferred to a nitrocellulose membrane using a
semidry transfer cell. Next, Tris buffered saline Tween
(TBST) containing 5% nonfat milk was added for
overnight incubation at 4℃ to block nonspecific
binding. After blocking, membranes were incubated
with primary antibodies against Bcl-2, Bax, Fas,
Caspase-9 and Caspase-3 and β-Actin for 2 h, and the
membrane was rinsed with PBS containing 0.05%
Tween 20 and filtered three times. The secondary
antibody was added, followed by incubation for 1 h
and washing with TBST three times. The membrane
was placed into DAB substrate solution and reacted for
3 min. The reaction was stopped by adding water, and
then an imager was used for radiography.
Statistical analysis
Statistical analysis was conducted with SPSS17.0
statistical software (SPSS, Inc., Chicago, IL, USA).
Data from each group were expressed as the average
value ± standard deviation and then evaluated by
one-way ANOVA. P < 0.05 indicated that the results
were statistically significant
Results
Inhibition effect of TSRP on transplanted tumor of
nude mice bearing lymphoma cells
After intragastric administration with different
concentrations of TSRP for 3 weeks, the weights of the
excised tumors in the low-dose and high-dose TSRP
groups were 0.86 ± 0.01 and 0.21 ± 0.03 g,
respectively, and were obviously lower in mass
compared to those from the control group (1.34 ± 0.50
g). Based on the formula for determining the tumor
inhibition rate: tumor inhibition rate = (1 － tumor
weight of the treatment group/tumor weight of the
control group) × %. The tumor inhibition rates for the
high-dose and low-dose TSRP groups in mice
transplanted with tumors were 90.84% and 27.96%,
respectively. These values were significantly different
compared to the control group (Both P < 0.001, Table
1).
Change in size of transplanted tumor after
treatment with TSRP
As the intragastric administration time increased,

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tumor size changed in the low-dose and high-dose
TSRP groups compared to those in the control group,
showing significant differences. This reveals that
TSRP strongly inhibits the growth of Raji cells in vivo
(Table 2).
TUNEL analysis of apoptosis of transplanted tumor
cells
Apoptotic cells were stained for fluorescence analysis
under a laser confocal microscope (×200). The
high-dose group contained more apoptotic cells than
the control group, and apoptotic cells presented a
central distribution. Using Leica Confocal software,
the fluorescence intensity of the three groups of
lymphoma apoptotic cells was quantitatively analyzed.
The results showed that the fluorescence intensity of
the control group was 10.49 ± 2.32, while these values
in the 20 and 100 mg/kg TSRP groups were 26.14 ±
2.42 and 82.43 ± 7.81, respectively. Compared to the
control group, these differences were significant
(Both P < 0.001, Figure 1).
Expression of Bcl-2, Bax, and Fas of transplanted
tumor determine by immunohistochemistry
analysis
The results showed that Bcl-2 expression of Raji cells
from the tumor treated with TSRP was clearly reduced
compared to in the control group, while the Bax
expression was clearly increased compared to that in
the control group. This indicates that TSRP induces
Raji cell apoptosis by reducing Bcl-2 expression and
increasing Bax expression. Additionally, Fas
expression in cells treated with TSRP was clearly
reduced compared to that in the control group,
demonstrating that TSRP induces Raji cell apoptosis
also by reducing Fas expression (Figure 2).
Inhibitory effect of TSRP on Raji cell proliferation
with MTT analysis
MTT analysis was conducted to determine whether
100-400 µg TSRP inhibited Raji cell growth
dose-dependently. At a concentration of 400 μg/mL,
the absorbance was 0.366 ± 0.031 compared to 0.780 ±
0.052 in the control group (P < 0.001 Table 3).
Table 1 Comparison of the tumor weight, size and tumor inhibition rate of each group after intragastric
administration with TSRP
Groups Tumor weight (g) Size (mm3
) Tumor inhibition rate (%)
Control 1.34 ± 0.50 2807.70 ± 128.43 -
TSRP (20 mg/kg) 0.86 ± 0.01 2022.59 ± 82.39#
27.96
TSRP (100 mg/kg) 0.21 ± 0.03*
257.29 ± 43.65#
90.84#
Compared with the control group: #, P < 0.001; *, P = 0.007.
TSRP, Total saponins of Rubus parvifolius L..
Table 2 Tumor size change of the nude mice after intragastric administration (mm3
)
Groups Intragastric administration time (day)
8 11 14 17 20
Control 307.46 ± 13.83 456.57 ± 15.98 1093.29 ± 125.19 2080.01 ± 112.03 2807.70 ± 128.43
TSRP
(20mg/kg)
212.51 ± 18.31 430.76 ± 10.28 998.43 ± 90.76 1122.59 ± 132.39#
2022 ± 182.39#
TSRP
(100mg/kg)
207.22 ± 4.84 214.34 ± 8.32 235.45 ± 7.98# 243.15 ± 23.65# 257.29 ± 43.65#
#, compared with the control group P < 0.001.

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Control 20mg/kg TSRP 100mg/kg TSRP
Figure 1 Comparing Raji cell apoptosis of the transplanted tumor of each group with TUNEL analysis
TSRP, Total saponins of Rubus parvifolius L.. #, compared with the control group, P < 0.001.
Bcl-2
Bax
Fas
Control 20mg/kg TSRP 100mg/kg TSRP
Figure 2 Expression of Bcl-2, Bax and Fas of the transplanted tumor of each group (×200)

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TSRP induction of Raji cell apoptosis observed by
DAPI staining
The nucleus of the control group had a regular and
round shape and showed uniform staining. As the
TSRP concentration increased, the nucleus became
cracked to form round bodies of different sizes
wrapped by the cell membrane (known as apoptotic
bodies). Apoptosis was clearly observed (Figure 3).
Expression of Bcl-2, Bax, Fas, Caspase-9, and
Caspase-3 evaluated by western blot analysis
Addition of TSRP significantly inhibited Bcl-2 activity
and increased Bax activity in Raji cells in a
dose-dependent manner. This revealed that TSRP
induces apoptosis by inhibiting Bcl-2 and increasing
Bax expression. TSRP also inhibited Fas expression in
a dose-dependent manner. Regarding the pathway by
which TSRP induces Raji cell apoptosis in vitro, we
found that Caspase-9 and Caspase-3 expression was
also increased with increasing TSRP concentrations
(Figure 4, Table 4). This suggests that TSRP induces
Raji cell apoptosis through the chondriosome pathway.
Table 3 Effects of TSRP at different concentrations with MTT
Groups Absorbance Inhibition rate (%)
Control 0.780 ± 0.052 -
TSRP (100μg/ml) 0.639 ± 0.024 18.15 ± 3.33
TSRP (200μg/ml) 0.531 ± 0.034 32.21 ± 4.89
TSRP (400μg/ml) 0.366 ± 0.031 53.46 ± 4.90#
#, compared with the control group, P < 0.001. TSRP, Total saponins of Rubus parvifolius L..
Control 100μg/mL TSRP
200μg/mL TSRP 400μg/mL TSRP
Figure 3 Morphology comparison after TSRP induces Raji cell apoptosis with DAPI analysis (fluorescence
microscope × 200).

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Discussion
Presently, lymphoma is treated by traditional
chemotherapy, targeted drug combined with
chemotherapy, or radiotherapy [5-8]. However, these
treatments have numerous serious side effects such as
immune system and hemopoietic system damage. Thus,
TCM may be valuable for inhibiting lymphoma cell
proliferation and inducing the apoptosis of these cells.
Herba RP is a plant in the genus Rubus L. and has
fairly standard features of the genus; its components
can remove pathogenic heat from the blood, arrest
bleeding, remove obstructions and relieve pain, induce
diuresis, and reduce edema, among other effects.
Previous studies have mainly focused on
cardiovascular effects of this plant, but its effects on
tumors remain unclear. In preliminary studies, we
showed that [2] TSRP significantly induced K562 cell
apoptosis in vivo and in vitro, and the mechanism was
related to increased expression of PARP, Caspase-3,
and Caspase-9 and inhibition of the expression of
Bcl-2. We also found that TSRP clearly inhibited the
phosphorylation level of STATS in vitro and increased
the phosphorylation levels of AMPK and JNK. A
high-dose TSRP can significantly inhibit the
phosphorylation level of Akt and mTOR. Therefore, it
is necessary to explore the anti-lymphoma effects of
TSRP. In this study, we found that administration of
TSRP (at concentrations of 20 and 100 mg/kg) to nude
mice inhibited the transplanted tumor. In vitro, an MTT
assay showed that 100-400 μg/mL TSRP had
inhibitory effects on Raji cells in a dose-dependent
manner. This reveals that TSRP has fairly strong
inhibitory effects on lymphoma cell growth in vivo and
in vitro.
The cell death process triggered by the in vivo and
in vitro factors in the cell is known as cell apoptosis.
Alterations in the normal apoptosis procedure in the
organism can result in many diseases such as tumors
[9]. In vivo, the TUNEL method showed that apoptotic
cells also increased with increasing TSRP
concentrations. Compared to the control group, the
difference was significant. Therefore, TSRP in vivo
inhibits lymphoma cell growth through apoptosis.
DAPI analysis suggested that the cell nucleus was
cracked to result in the formation of apoptotic bodies
of different sizes as the TSRP concentration increased
in vitro, showing obvious apoptosis. Therefore, TSRP
clearly induced Raji cell apoptosis in vivo and in vitro.
The endogenous apoptosis pathway (chondriosome
pathway) originates from pressure inside of a cell. For
example, rays, toxic drugs, a lack of growth factors,
and other factors result in Caspase-9 activation, trigger
the caspase cascade reaction, and cause cell apoptosis
[10-12]. Caspase-3 is the most important molecule
involved in apoptosis in the caspase family, and it is
related to morphology changes such as chromatin
agglutination, karyopyknosis, and karyorrhexis during
apoptosis [13]. In early stages of apoptosis, Caspase-3
receives a signal from upstream caspase molecules and
becomes activated; the relevant cytoplasm and karyon
substrate is disrupted to result in cell apoptosis [14-15].
According to western blot analysis, the expression of
Caspase-9 and Caspase-3 also increased with
increasing TSRP concentrations in a dose-dependent
manner. This indicates that TSRP in vitro induces
apoptosis through the chondriosome pathway. Further
in vivo experiments are required to confirm these
results.
Bcl-2 and Bax are two important members of the
Bcl-2
Bax
Fas
Caspase-9
Caspase-3
β-Actin
Control 100 200 400
μg/mL TSRP
Figure 4 Effect of TSRP at different concentrations on activity of Raji cell apoptosis controlling proteins
Bcl-2, Bax, Fas, Casepase 9 and Casepase 3
Compared with the control group: #, P < 0.001; *, P = 0.027. TSRP, Total saponins of Rubus parvifolius L..

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Bcl-2 gene family and have contrasting functions:
inhibiting cell apoptosis and promoting cell apoptosis,
respectively. The formation of Bax homodimers
induces apoptosis. Bcl-2 protein inhibits apoptosis
mainly by combining with Bax to form heterodimers
[16-18]. The ratio of Bcl-2 to Bax also determines the
formation of such dimmers to finely control cell fate
[19-22]. The Fas/CD95 system is a signal transduction
system that mediates cell apoptosis and has been
widely studied; this system belongs to the family of the
tumor necrosis factor receptor and nerve growth factor
receptor, which mediates cell death and is known as
the death domain; it activates the Fas death signal and
causes the tumor cell to undergo apoptosis [23-24].
Our results showed that the rate of positive Bcl-2
expression in cells treated with TSRP in vivo was
obviously reduced compared to that in the control
group. Additionally, the Bax expression was obviously
increased compared to in the control group. This
reveals that TSRP induces Raji cell apoptosis by
reducing Bcl-2 expression and increasing Bax
expression. We also found that Fas-positive expression
in cells treated with TSRP was clearly reduced
compared to that in the control group, suggesting that
TSRP induces Raji cell apoptosis by reducing Fas
expression. In vitro experiments showed the same
results. Therefore, TSRP induces Raji cell apoptosis
likely by altering the Bcl-2/Bax ratio and Fas signal
pathway.
It remains unclear whether Bcl-2 affects apoptosis
controlled by the Fas system. In some cells, Bcl-2 can
inhibit CD95 and induce apoptosis. However, in other
cells, there is no correlation between Bcl-2 and CD95.
This will be examined in our future studies. Our results
revealed a correlation between Bcl-2 and Fas
expression.
Conclusion
TSRP induces Raji cell apoptosis by inhibiting Bcl-2,
increasing Bax expression, and reducing Fas
expression in vivo and in vitro. Additionally, TSRP has
some effects on the chondriosome pathway of
apoptosis in vitro.
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