In Vivo and In Vitro Study on the Mechanism of Anticervical Cancer Effects of Corilagin in Mice

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Introduction
Cervical cancer is a malignant tumor that occurs at the junction of squamous epithelial cells in cervical vagina.It can also infect the transition zone and columnar epithelial cells in the inner lining of the neck tube.Globally, cervical cancer is the fourth most common cancer and the fourth leading causes of cancer deaths among women.In 2018, there were 570,000 patients with cervical cancer and 311,000 reported deaths from the cancer in the world [1][2][3].Surgical operation together with radiotherapy and chemotherapy are the common methods of cervical cancer treatment.However, the strategies do not efectively eradicate the tumor tissue in patients with locally advanced, recurrent, and metastatic cervical cancer.In terms of chemotherapeutic drug selection, 5-fuorouracil (5-FU) has been used for many years as an anticervical cancer drug for optimal postoperative survival, but recently frequent emergence of drug resistance has limited the clinical use of 5-FU [4], and there is an urgent need for treatment methods that are both effective and low toxic.Tis shall improve the outcome of cervical cancer treatment and reduce the mortality rate of patients with the cancer.
Corilagin is a type of ellagitannin which is a natural component in several ethnopharmacological plants.Its chemical formula and molecular weight are C 27 H 22 O 18 and 634.45, respectively.It is an of-white needle-like crystalline powder which is soluble in methanol, ethanol, acetone, and dimethyl sulfoxide.To date, Corilagin has been detected in 50 species of 16 plants families [5].Te ellagitannin is one of the main active ingredients of various ethnic medicines, such as Phyllanthus urinaria [6], matsumura leaf fower herb [7], and Euryale ferox Salisb.seed [8].Recent pharmacological studies have found that Corilagin has antitumor [9], antiinfammatory [10], antibacterial [11], and antiviral [6] effects.It has signifcant application prospects especially in the antitumor pharmacological aspect.Currently, there is no in vivo study of Corilagin against cervical cancer.
Results of our preliminary studies found that Corilagin isolated from the shell of the Euryale ferox Salisb.seed has a signifcant inhibitory efect on proliferation, migration, and invasion of cervical cancer HeLa cells.Tis indicates that ellagitannin has enormous potential in combating cancer [12].Terefore, the current study started from investigation of the chemical structure of Corilagin.Te study later applied the network pharmacology technology to predict its antitumor target genes and their mechanisms.Experiments were conducted on the efects of Corilagin on U14 cell proliferation, cell cycle, and apoptosis.Finally, its antitumor efect and molecular mechanism were studied by establishing a subcutaneous xenograft model of cervical cancer in mice to investigate the efect of Corilagin on the growth of xenograft.

Prediction of Potential Target Genes for Anticervical
Cancer Efect of Corilagin.First, the secondary chemical structure of Corilagin was searched in the TCMSP database.Te PharmMapper and SwissTargetPrediction were then used to predict the target that can bind Corilagin.Te related genes of cervical cancer were screened using GeneCards, OMIM, and DisGeNET databases to establish a cervical cancer target database.Te intersection with the above targets was the potential target of Corilagin's anticervical cancer.

Construction of Potential Target Interaction Networks for the Anticervical Cancer Efect of Corilagin.
Te potential targets obtained in Section 2.2 of this study were used to construct the PPI network map using the STRING database.Te targets that did not interact with other proteins were deleted and the derived data were used to make a protein interaction map using Cytoscape_v3.6.1.

Establishment of a Pathway Network for the Potential
Anticervical Cancer Target of Corilagin.Te targets obtained in Section 2.2 were uploaded in the DAVID database.Te KEGG pathway enrichment analysis was performed to investigate the anticervical cancer targets of Corilagin.

Detection of Cell Proliferation Inhibition
Rate.Te cells in the logarithmic growth phase were counted and laid according to 3,000 cells/well.Te concentration of Corilagin and 5-FU was set at 0, 200, 400, 600, 800, and 1000 μmol/L.Te treatment time were set at 24 h and 48 h.Te OD values were then assayed according to the growth manipulation instructions provided with the CCK-8 kit, and the cell proliferation inhibition rate was calculated according to formula (1).Te ODtest group represents the OD of the group with cells but without drug administration; the ODblankgroup represents the OD of the drug-free and cellfree group; the ODcontrol group represents the OD of the administered group.

Organ Coefcients' Calculation and H&E Staining
Analysis.Te livers and kidneys of mice were dissected, weighed to calculate organ coefcients, and organs were sectioned and stained with H&E.Tey were then examined microscopically to see if the drug had damaged their tissues.

ELISA: Detection of IL-2, TNF-α, and IL-10 Indicators in
Serum.After 30 min of indoor coagulation, the levels of IL-2, TNF-α and IL-10 in the serum were detected through centrifugation at 1000 g for 10 min according to the procedure described on the ELISA kit.

RT-PCR: Detection of the mRNA Expression.
Based on the results of the previous experiments, the group with the best results was selected for the experiment.Te RNA of the tumor tissue was extracted through the Trizol method and reverse transcription was then performed.Te cDNA obtained was stained with SYBR and then subjected to the realtime fuorescence quantitative analysis through a two-step procedure.Te amplifcation procedure included an initial incubation at 60 °C for 30 s, followed by incubation at 95 °C for 3 s, and then 40 cycles in a sequence.
2.12.Western Blot Detection of Protein Expression.Te tissues were collected from the high-dose group and the model group.Te weighed tumor tissue was placed in EP tubes and their protein was quantifed using the BCA kit.Protein supernatant was prepared with bufer at the ratio of 4 : 1(bufer: protein supernatant) and then added to the protein sample.Te protein can be stored in the chain form for 10 min at temperature between 80 and 95 °C.Te protein samples were injected into the sample pool of the gel preparation instrument and then added into the separation gel sample wells for separation through electrophoresis.Te gels were cut according to the position of protein markers, and the membranes were transferred at 100 V for 90 min in an ice-cold water bath.Te membrane was cleaned thrice with TBST for 5 min each time.Te frst antibody was added and incubated overnight at 4 °C.Te primary antibody was washed three times with TBST.Te second antibody was then added and incubated at room temperature for about 1 h.Te second antibody was fnally washed three times with TBST.After adding the developer, the images were scanned using a gel imager and analyzed using the Image J analysis software.

Statistical Analysis.
GraphPad Prism 8.0.1 was used to plot graph for the obtained data and analyzed the signifcant diference.Te results are expressed as mean ± standard deviation (mean ± SD) and the diference between groups was tested using t tests.Statistically signifcant diference was set as p < 0.05.

Potential Target Genes for the Anticervical Cancer of
Corilagin.Trough the drug structure (Figure 1(a)), the present study used the Uniprot database to screen out 334 target genes that can be combined with Corilagin.A total of 7,076 genes related to cervical cancer were screened using GeneCards, OMIM, and DisGeNET databases.Both databases (OMIM and DisGeNET) drew the Venn diagram as presented in Figure 1(b), which shows that there were 222 common target genes.

Protein Interaction Network Analysis.
Te protein interaction data of the potential target genes were obtained from the STRING database and the data were imported into Cytoscape to construct the "protein interaction" network diagram (Figure 2).Te color was set based on the degree value.Te larger the degree value, the darker the color and the higher the correlation with other targets; MAPK1, SRC, PIK3R1, TP53, HSP90AA1, HRAS, RHOA, and other important genes were screened.
Te 31 key targets with high degree values were screened to be visible in the tumor signaling pathway by the KEGG Mapper function in the KEGG database.According to Figure S1, we found that the selected targets include BCL-2, BCL XL, CASPASE3, and other related apoptotic targets.Furthermore, it was noted that the selected targets mainly function through PI3K/AKT pathway and MAPK signaling pathway.Tis indicates that Corilagin may afect the PI3K/ AKT and MAPK signaling pathways to cause tumor apoptosis and achieve the anticervical cancer efect.

KEGG Enrichment Pathway Analysis.
Te KEGG enrichment analysis was carried out on the relevant target genes using the DAVID database.A total of 105 signifcantly enriched signal pathways were screened out, broad pathways were excluded, and 17 more signifcant pathways were fnally obtained (Figure 3).It was evident that the pathways with larger enrichment factors and a larger number of counts under the pathway include PI3K-AKT, RAS, RAP1, MAPK, FOXO, and HIF-1 signaling pathways.
RAS protein is a type of small GTPases superfamily which is composed of KRAS, HRAS, and NRAS.It plays a regulatory role in the growth of many normal cells.Missense mutations in the RAS gene have been found in 25% of human cancers.Te gene makes the research on the treatment of cancer by inhibiting the RAS gene become a hot spot.RAS binds to a variety of efector proteins and hence can transmit downstream signals to control cell proliferation, diferentiation, adhesion, migration, and survival.Research studies on the targeted RAS focuses on inhibition of downstream signaling pathways because it is difcult to directly target the small molecular mutant RAS.Te RAS sends signals through a variety of efectors.Te signals mainly stimulate both the MAPK and PI3K/AKT pathways [13].Furthermore, RAP1 has a high degree of sequence similarity with RAS and it is a member of the RAS family of small GTPases.Te RAP1 has overlapping binding partners.
Mechanistically, RAS and RAP1 cooperate to initiate and maintain ERK signaling.RAP1 can also be directly combined with the RAS binding domain that stimulates PI3K [14].Tis shows that both RAS and RAP1 signaling pathways can play a signifcant role in infuencing the MAPK and PI3K/AKT pathways.In conclusion, it was evident that the mechanism of Corilagin against cervical cancer may be related to PI3K/AKT and MAPK signaling pathways.

Inhibition of Cell Proliferation in U14 Cervical Cancer.
Te efect of Corilagin on proliferation of U14 cells was assessed after Corilagin was allowed to act on the cells for 24 h or 48 h.Results of the analysis are as presented in Figure 4.It was evident that Corilagin has a signifcant inhibitory efect on the proliferation of U14 cells.It was found that Corilagin at a concentration of 200 μmol/L showed proliferation inhibition of U14 cells after 24 h or 48 h, with inhibition rates of 1.51% and 1.64%, respectively, which were lower than that of the positive drug (5-FU).
At the concentration of 400 μmol/L, Corilagin showed a higher inhibitory efect as compared with that at concentration of 200 μmol/L.Te inhibition rate of Corilagin (400 μmol/L) on U14 cell proliferation at 24 h and 48 h was 64.72 ± 6.05% and 75.18 ± 7.74%, respectively, which were higher than the inhibitory efect of 5-FU (positive control group).Te inhibitory efect of Corilagin at concentrations higher than 400 μmol/L was higher than that of 5-FU.Furthermore, its IC 50 value after 24 h and 48 h exposure to the U14 cell was 372.4 μmol/L and 394.5 μmol/L, respectively (Supplement materials Table S2).

Efects of Corilagin on U14 Cell Cycle.
Results of the present study showed that the number of cells in the G0/G1 phase remained basically unchanged as compared with that in the model group after Corilagin was applied to U14 cells for 48 h, at concentrations lower than 400 μmol/L (Figure 5).Furthermore, it was noted that the number of cells in S phase increased, whereas that in the G2/M phase was reduced as compared with that in the control group after drug exposure to the cells for 48 h at drug concentrations lower than 400 μmol/L.
On the other hand, when the concentration of Corilagin was higher than 400 μmol/L, it was found that the number of cells in the G0/G1 phase increased in concentrationdependent manner.However, the number of cells both in the S and G2/M phases signifcantly decreased as compared with the control (model) group.Tis showed that reduced concentrations of Corilagin (below 400 μmol/L) blocked proliferation of U14 cells in the S phase, whereas higher concentration (above 400 μmol/L) blocked proliferation of the cells in the G0/G1 phase.
Progression of the cell cycle is maintained by a variety of molecules such as growth factors and hormones.It also involves a series of strictly regulated molecular processes that ultimately causes cell division.Te cell cycle includes four stages: G1 (premitosis), S (DNA replication), G2 (mitosis preparation), and M (genetic material and

Efects of Corilagin on Apoptosis of U14 Cells.
Results of the current study showed that there was an increase in the rate of apoptosis with increase in the concentration of Corilagin as compared with the rate in the control group (Figure 6).For instance, when the drug concentration was 400 μmol/L, the rate of apoptosis was signifcantly diferent, whereby the rates for early and late withering cells were 13.6 ± 3.89% and 35.22 ± 13.69%, respectively.In summary, it was evident that Corilagin has a signifcant efect on the apoptosis process of the U14 cells.

Efects of Corilagin on the Growth of Transplanted Tumors
and Tumor Tissue Apoptosis in Mice.It was found that both the Corilagin and positive drug (5-FU) groups had diferent degrees of inhibition on the growth of transplanted tumors in mice (Figure 7).Te rate of tumor inhibition by Corilagin in the low-dose group was 43.39%, whereas the tumor volume was reduced by 51.92% as compared with the negative control group.Furthermore, the inhibition efect of Corilagin and 5-FU drugs in the high-dose group was more signifcant, and the tumor inhibition rates were 53.99% and  Journal of Food Biochemistry 54.30%, respectively (p < 0.05), whereas the tumor volumes were decreased by 62.92% and 56.70%, respectively (p < 0.01) (Supplement Materials Table S3).Tis study showed that the tumor tissues in the negative control were mainly normal tumor cells with a small amount of apoptotic cell particles (Figure 8).However, it was evident that both the high-dose Corilagin and the positive drug (5-FU) groups contained a large area of apoptotic cells.Te apoptotic area observed in the high-dose Corilagin group was larger than that of the 5-FU group which indicated that Corilagin could more signifcantly induce tumor tissue apoptosis as compared with the positive drug.

Te Efect of Corilagin on Organ Coefcient and the
Histopathology of Mice. Figure 9 shows that the kidney organ coefcients of the model group were comparable with those in the blank group, but the liver organ coefcients (p < 0.05) were higher in the model group than in the blank group.Tis indicates that the liver function of the transplanted tumor mice may decrease and the kidney tissue may not be afected.Notably, between the model group and the drug administration groups, there was no signifcant difference in the kidney organ coefcients, indicating that neither Corilagin nor 5-FU had a signifcant efect on the kidney tissue.Te organ coefcient of the liver in mice treated with high doses of Corilagin decreased signifcantly (p < 0.05), which indicates that high-dose Corilagin may confer protection on the liver function.Morphological and structural analyses showed that liver cells in the blank group were normal, and the nucleus as well as the cytoplasm were evenly distributed (Figure 10).Except for the blank group, a large amount of hepatocyte edema was seen in the liver tissues of all other groups.Tis was accompanied by a loose and slightly stained (black arrow) cytoplasm.Neutrophil infltration (red arrow) was seen around the bile duct in the portal area.Moreover, a few extramedullary hematopoietic foci (yellow arrows) were seen indicating that the liver of the transplanted tumor mice had lesions.
Te surface envelope of the kidney tissue is composed of a dense connective tissue with uniform thickness.Te kidney parenchyma consists of superfcial cortex and deep medulla.According to the pathological observation of kidneys (Figure 11), the cortical medulla of the kidney tissues were clearly demarcated in all groups, the glomeruli were evenly distributed in the cortex, and the number of cells and matrix in the glomeruli were uniform.Te renal tubule epithelial cells were round and full, the brush borders were neatly arranged, and the medulla is not signifcantly damaged.Te connective tissue between the urinary tubules forms the interstitial fuid.In this study, the interstitial did not proliferate; and no obvious infammatory changes were seen.
In summary, the liver of mice transplanted with U14 cells showed severe damage compared with normal mice.Te high-dose Corilagin and 5-FU did not signifcantly alleviate liver damage.Moreover, there was no signifcant diference in the kidney tissues of the mice across the groups, which indicated that Corilagin and 5-FU had no signifcant toxic efects on the kidneys, and the results were consistent with the results of organ coefcients.In practice, the liver and kidney functions are jointly monitored, and drugs targeting the liver and kidneys are similar.

Te Efect of Corilagin on IL-2, TNF-α, and IL-10 on
Serum Levels in Mice.As shown in Figure 12, mice in the model group had signifcantly lower serum levels of IL-2 and higher levels of IL-10 than those in the blank group.However, there was no signifcant diference in the content of TNF-α between the two groups.Te levels of IL-2 and TNF-α in the 8 Journal of Food Biochemistry high-dose Corilagin group and the positive drug group were signifcantly higher, whereas the content of IL-10 was signifcantly lower (p < 0.05).

Discussion
Studies have shown that Corilagin has an inhibitory efect on a variety of cancers.Attar et al. [17] showed that Corilagin was antigastric cancer in vitro by inducing apoptosis, autophagy, and increasing reactive oxygen species generation; Wu et al. [18] isolated Corilagin from Phyllanthi Fructus and confrmed that it was antiesophageal cancer by activating mitochondrial and endoplasmic reticulum stress signaling pathways.Te main purpose of this study was to verify the inhibitory efect of Corilagin on cervical cancer and to investigate its mechanism through in vitro and in vivo experiments.
Results of the CCK-8 assay in the present study showed that Corilagin signifcantly inhibited the proliferation of cervical cancer U14 cells of mice.Furthermore, it was found that Corilagin afected the cell growth by afecting the cell cycle distribution of U14 cells.Terefore, it was evident that Corilagin can signifcantly inhibit the growth of U14 tumorbearing mice.Te rate of tumor inhibition in the high-dose Corilagin group (30 mg/kg) was similar to that of the positive drug 5-FU (20 mg/kg).Te results indicated that Corilagin could signifcantly inhibit the proliferation of the cervical cancer tissue.Organ coefcient and pathological section methods were used to detect drug toxicity of 10 Journal of Food Biochemistry Corilagin in mice.In toxicology research, the organ gravimetric analysis is considered to a vital indicator to detect whether a compound is harmful to the human body.Differences in organ weight are often compared with diferences in body weight between diferent treatment groups, and the weight error can hence be eliminated by introducing organ coefcients [19].Studies have shown that the coefcients of the liver, kidney, and spleen organs play a crucial reference role in prediction of a compound toxicity in the body [20].In the present study, it was found that the organ coefcient was greater than that of the normal group, indicating that there were changes in the organ edema and congestion.In addition, it was evident that the organ coefcient was lower than that of the normal group, indicating organ atrophy or degenerative changes.Results of this study showed that Corilagin can signifcantly reduce the liver organ coefcient, but has no signifcant efect on the kidney organ coefcient.Tis indicated that Corilagin has a protective efect on the liver function, but may not have a signifcant toxic efect on the kidney tissues of mice.Furthermore, the results were basically consistent with those in the pathological sections of this study.Subsequently, the mechanism of the anticervical cancer inhibits of Corilagin was studied.Te results of network pharmacology indicated that Corilagin may achieve the efect of anticervical cancer by afecting PI3K/AKT, MAPK, and other signaling pathways.Furthermore, its mechanism of action may be associated with the signal pathway of apoptosis.Flow cytometry and TUNEL staining confrmed that Corilagin can induce apoptosis to afect the growth of the cervical cancer tissue.
Te efects of Corilagin on mouse serum biochemical indices were studied through ELISA.It was found that helper T cells (T cells) play a vital role in immune response of the body.T cells have two subgroups: T1 and T2.Tumor progression is often accompanied by T1/T2 metastasis to T2 [21], which is mainly because the imbalance of T1/T2 enables local T cells to escape the immune response to tumor tissues.T1 cells mainly secrete cytokines such as TNF-β, TNF-α, and IL-2 which play a cytotoxic efect on killing cells.Te IL-2 can not only stimulate NK cells to produce interferon, but also strengthen the function of T1 cells and hence can strengthen the immune system of the body and induce tumor immune response [22].TNF-α, or tumor necrosis factor-α, is not only a crucial regulator of the infammatory response in the body, but can also form a complex with cell surface receptors or increase the adhesion of neutrophils to endothelial cells.
TNF-α has a direct cytotoxic efect on tumor cells [23].T2 cells mainly secrete cytokines such as IL-6 and IL-10.Te cytokines IL-10 can reduce the proliferation of T cells, inhibit the antitumor immune response and the phagocytosis of monocytes/macrophages, and afect the growth proliferation as well as apoptosis of tumor cells.It has been found that the upregulation of IL-10 is closely related to the deterioration of cervical cancer [24].Tis could be because the HPV oncogene protein E6 can afect the expression of IL-10 and the HPV escapes the detection of the immune system, which causes the occurrence of cervical cancer after continuous HPV infection [25].
Te results of the current study showed that Corilagin can signifcantly increase the level of IL-2 and TNF-α in the  serum of mice.Furthermore, treatment with Corilagin reduced the level of IL-10, increased the level of T1 cytokines, and reduced the level of T2 cytokines.Te fnding indicates that Corilagin may enhance the antitumor immune response of the body by restoring the balance of the T1/T2 cell response and hence delay the progression of cervical cancer.
RT-PCR and Western blot analysis were used to validate the expression of target genes in network pharmacology prediction pathways.EGFR is an epidermal growth factor receptor that plays a signifcant role in growth and diferentiation of tumors.Terefore, the production of a large number of cancer cells accompanied by high activation, overexpression, or signaling imbalance of EGFR is an upstream target of most tumor-related pathways.When EGFR is activated, EGFR ligands bind to the EGFR receptors, and the two undergo dimerization to stimulate RAS proteins in the downstream of the EGFR signaling pathway.Tese cause phosphorylation cascades, activate PI3K and MAPK signaling pathways, and afect the occurrence as well as development of tumors [26,27].A key downstream mediator of the PI3K signaling cascade is the serine/threonine kinase (AKT).Tis phosphorylates a variety of intracellular proteins to promote progression and activity of the cell cycle [28].Te MAPK subfamily includes ERK, JNK, and P38 that play key roles in the survival and apoptosis of various cancer cells through specifc phosphorylation.Fluorescence quantitative results of the present study showed that after Corilagin treatment, the mRNA expressions of EGFR, and the RAS-related genes HRAS, AKT, and ERK were significantly decreased, whereas the mRNA expressions of JNK and P38 genes were increased after the treatment.Results of the Western blot assays showed that the expression levels of EGFR and p-Akt (Tr308) were signifcantly decreased, and the expression level of JNK was also signifcantly increased following the treatment of cells with Corilagin.Tis shows that Corilagin can trigger EGFR/RAS/AKT and EGFR/RAS/ MAPK signaling pathways.
Tere are two pathways of apoptosis that are induced by complex cytotoxicity: death receptor pathway and mitochondrial pathway.Te death receptor pathway mainly occurs through the binding of trimeric Fas to the receptor cytosolic domain and thereby triggering the Caspase cascade.Furthermore, the pathway relies on the interaction of the proteins in the Bcl-2 family to transport cytochrome C into the cytoplasm.After binding to apoptotic protease activator I, the Caspase-9 and Caspase-3 are activated, triggering a cascade reaction that causes apoptosis [29].Terefore, it is evident that Bcl-2 family genes play a key role in the process of apoptosis.Antiapoptotic proteins of Bcl-2 family include BCL-2 and BCL-XL, whereas the proapoptotic proteins include Bad and Bim [30].
Te fndings of RT-PCR showed that Corilagin treatment signifcantly decreased Bcl-2 and signifcantly increased the mRNA expressions of Caspase3, Caspase9, Cyto C, and Bim.Results of the Western blot revealed that after treatment with Corilagin, the Caspase9 was signifcantly increased and this was an indication that Corilagin can induce cell apoptosis.
FOXO transcription factor is a downstream target for PI3K/AKT.AKT kinase-mediated phosphorylation regulates the transcriptional activity, whereas the cell activity is infuenced by the transcriptional regulation of death receptor ligand and expression of Bim [31].Te phosphorylation of FOXO3A promotes the expression of downstream target genes, such as Bim, and the BH3 domain of Bim initiates the apoptotic cascade by binding BCL-2 and BCL-XL [32].Furthermore, previous studies have found that tumor cells can induce apoptosis through AKT/FOXO3A/Bim signaling pathway [33].Te results on the levels of mRNA showed that the mRNA expression of FOXO and Bim was signifcantly upregulated.Tis suggests that Corilagin can induce apoptosis by activating the FOXO signaling pathway.Besides, this fnding was same as the result that Corilagin promoted apoptosis of U14 cells in vitro.
In conclusion, it was evident that Corilagin can significantly upregulate mRNA expression of genes Bim, Cas-pase3, Caspase9, and Cyto C as well as Caspase9 protein expression.In addition, it was found that Corilagin can signifcantly downregulate the mRNA level of gene Bcl-2 to activate AKT/FOXO3/Bim and JNK/P38 MAPK signaling pathways.Lastly, it was evident that the inhibition of AKT/ HIF signaling pathway induces apoptosis in tumor tissues.

Conclusion
Network pharmacology can more accurately predict the anticervical cancer mechanism of Corilagin.Te present study evidently showed that Corilagin can signifcantly inhibit the proliferation of U14 cells and afect the cell cycle and apoptosis in vitro.Results of in vivo experiments in the current study showed that Corilagin can signifcantly inhibit tumor growth, without causing signifcant toxicity to the body, enhance antitumor immune response, and induce tumor tissue apoptosis to achieve the efect of anticervical cancer in mice.However, there are no in-depth in vivo studies on the absorption, metabolism, and bioavailability of Corilagin and the pharmacokinetic process of Corilagin is still unclear.Terefore, there is a need for more in-depth research studies on the dosage, form, and pharmacokinetic process of Corilagin in animal bodies.

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Journal of Food Biochemistry cytokinesis).Dysregulation of the cell cycle is the cause of abnormal cell proliferation and cancer[15].Previous studies have shown that Corilagin can afect the cell cycle in the tumor.Furthermore, the drug can block the G2/M phase of liver cancer cells and has been associated with the activation of P53-P21CIP1-CDC2/Cyclin B1[16].

Figure 2 :
Figure 2: Protein interaction network diagram of potential targets.Protein interaction network diagram: the higher the degree value, the darker the color.

Figure 9 :Figure 10 :
Figure 9: Te changes of organ coefcient in mice induced by Corilagin: (a) liver coefcient and (b) kidney coefcient.Te dose was 15 mg/kg in the low-dose group, 30 mg/kg in the high-dose group, and 20 mg/kg in the 5-FU group ( * p < 0.05 vs. 0 mg/kg).