Compound shougong powder inhibits the malignant phenotype of hepatocellular carcinoma cells by targeting the DNA damage repair pathway.

OBJECTIVES: Compound Shougong Powder (SGS), a traditional Chinese medicine formulation, has been used to treat cancer for many years with remarkable efficacy. However, the mechanisms underlying the therapeutic effect of SGS in Hepatocellular carcinoma (HCC) are not completely clear. METHODS: The survival and metastasis of HCC cells were examined by CCK-8 assay, EdU assay, Wound-healing and Transwell assay. The anti-tumour effect of SGS was studied using hoechst 33258 staining and flow cytometry. RNA sequencing was applied to detect the underlying mechanism. Comet DNA, qRT-PCR and WB experiments were performed for validation. In addition, HCC nude mouse model was constructed to detect SGS effect in vivo. KEY FINDINGS: SGS inhibited the proliferation, migration and invasion of HCC cells and induced apoptosis in vitro. In addition, SGS also suppressed tumour growth in a nude mouse model of HCC in a dose-dependent manner. RNA sequencing of the suitably treated HCC cells revealed significant changes in the expression levels of genes involved in the DNA damage repair pathway. The sequencing results were verified by Comet DNA, qRT-PCR, WB assays and molecular docking. CONCLUSIONS: Taken together, SGS inhibits the malignant phenotype of HCC cells by down-regulating DNA repair genes and consequently inducing DNA damage.

Effects of N6-Methyladenosine Modification on Cancer Progression: Molecular Mechanisms and Cancer Therapy.

N6-methyladenosine (m6A) is a major internal epigenetic modification in eukaryotic mRNA, which is dynamic and reversible. m6A is regulated by methylases ("writers") and demethylases ("erasers") and is recognized and processed by m6A-binding proteins ("readers"), which further regulate RNA transport, localization, translation, and degradation. It plays a role in promoting or suppressing tumors and has the potential to become a therapeutic target for malignant tumors. In this review, we focus on the mutual regulation of m6A and coding and non-coding RNAs and introduce the molecular mechanism of m6A methylation involved in regulation and its role in cancer treatment by taking common female malignant tumors as an example.

Roles of Major RNA Adenosine Modifications in Head and Neck Squamous Cell Carcinoma.

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer malignancy worldwide and is known to have poor prognosis. The pathogenesis behind the development of HNSCC is not fully understood. Modifications on RNA are involved in many pathophysiological processes, such as tumor development and inflammation. Adenosine-related RNA modifications have shown to be linked to cancer and may play a role in cancer occurrence and development. To date, there are at least 170 different chemical RNA modifications that modify coding and non-coding RNAs (ncRNAs). These modifications affect RNA stability and transcription efficiency. In this review, we focus on the current understanding of the four major RNA adenosine modifications (N6-Methyladenosine, N1-Methyladenosine, Alternative Polyadenylation Modification and A-to-I RNA editing) and their potential molecular mechanisms related to HNSCC development and progression. We also touch on how these RNA modifications affect treatment of HNSCCs.

Epithelial-mesenchymal transition classification of circulating tumor cells in lung and colon cancer patients: potential role in clinical practice.

BACKGROUND: Tumor cells undergoing epithelial-mesenchymal transition (EMT) display enhanced ability to enter the circulation, thereby being major source of circulating tumor cells (CTCs). In this study, we aimed to better understand the roles of CTC undergoing EMT in monitoring cancer progression. METHODS: We analyzed gene expression profiling of epithelial and mesenchymal markers in lung or colon tumor samples by mining TCGA database. We detected CTCs and classify their EMT phenotypes of 31 patients with lung or colon cancer by using a CanPatrol CTC-enrichment technique. RESULTS: The bioinformatic analysis indicated that mesenchymal markers were expressed in a subset of lung tumor samples, and its high expression was associated with poor survival of lung cancer patients. However, in colon cancer, majority of tumor samples expressed hybrid epithelial/mesenchymal markers. CTC analysis with EMT classification showed that the number of CTCs with mesenchymal phenotype was high in lung cancer patients with the advanced stage. Dynamic CTC analysis in a lung cancer patient indicated that CTC with mesenchymal phenotype was effective to monitor tumor progression. In a colon cancer patient, dynamic CTC analysis indicated that CTC with hybrid epithelial/mesenchymal phenotypes was an effective biomarker to guide therapy. CONCLUSIONS: Encouraging results from this proof-of-concept study show that CTC with mesenchymal phenotype or hybrid epithelial/mesenchymal phenotypes could be a potential biomarker for monitoring tumor progression in lung or colon cancer respectively.

[Pidotimod inhibits activation of latent Toxoplasma gondii infection induced by dexamethasone in mice].

OBJECTIVE: To study the inhibition effect of pidotimod (PT) on dexamethasone (Dem)-induced reactivated toxoplasmosis in mice. METHODS: A total of 96 female BALB/C mice were infected orally with 30 cysts of Toxoplasma gondii TgCtwh6 strain (genotype Chinese 1). 4 weeks later the mice were divided into three groups (A, B, and C). Mice of group A (Dem+NS) were given Dem [6 mg/(kg x d)] intraperitoneally and 200 microl normal saline given orally. Mice of group B (Dem+PT) were orally given pidotimod [100 mg/(kg x d)] and intraperitoneally injected with Dem[6 mg/(kg x d)] simultaneously. Each mouse in group C received 200 microl normal saline intraperitoneally. The mice were injected and given by gavage for 5 weeks. After treatment, three mice in each group were scarified weekly, and the survival time of the mice was recorded in days. Brain parasite burden and T. gondii DNA copies in serum were detected by quantitative real-time PCR. T cell subsets, cytokine profiles in each group were analyzed by flow cytometry, and CBA kit, respectively. RESULTS: On the second week after Dem administration, parasitemia appeared in group A; in 5 weeks 50% mice had parasitemia again, and 17 mice died. Comparatively, in group B parasitemia appeared on the third week after PT and Dem administration, in 5 weeks 25% mice had parasitemia again, and 7 mice died. Parasitemia did not appear in Group C. On the 21st day after Dem administration, T. gondii DNA copies in brain tissues of group A was (209 +/- 12) x 10(9), significantly higher than (62 +/- 10) x 10(9) in group B treated with PT (n = 3, P < 0.01). Flow cytometry test showed that on the 21st day after Dem administration, the proportions of Th1, Th2 and Treg cells in groups A and B were (4.0 +/- .5)% and (6.1 +/- 1.0)%, (0.6 +/- 0.1)% and (0.5 +/- 0.2)%, and (5.0 +/- 0.9)% and (7.0 +/- 1.2)%, respectively. There was significant difference in the percentages of Th1 and Treg between group B and A (P < 0.01). The levels of IFN-gamma, TNF-alpha in group A were (2.2 +/- 0.7) pg/ml and (20.1 +/- 5.0) pg/ml, respectively, lower than that of group B [(3.6 +/- 0.6) pg/ml and (32.0 +/- 8.0) pg/ml] (P < 0.01). No statistical significance was found in the levels of IL-4 and IL-10 between group A [(2.6 +/- 0.4) pg/ml, (39.0 +/- 6.0) pg/ml] and group B [(2.7 +/- 0.7) pg/ml, (40.0 +/- 8.0) pg/ml] (P > 0.05). CONCLUSION: Pidotimod can inhibit activation of latent Toxoplasma gondii infection induced by dexamethasone in mice. Th1 and Treg cells may contribute to the pidotimod/dexamethasone-induced immunoregulation.

[Kinetic distribution of cyst-forming Chinese isolate of Toxoplasma gondii in mice].

OBJECTIVE: To determine the kinetics of infection and cyst formation in CD1 mice following oral infection with cyst-forming Chinese isolate of Toxoplasma gondii TgCtwh1(genotype China 1, ToxoDB#9). METHODS: 50 CD1 female mice were obtained from specific pathogen-free (SPF) mouse colony in the Vital River Laboratories (VRL), Beijing. Mice were randomly divided into 10 groups each with 5 mice. All mice but control were peroral gavage infected with 50 cysts (1x10(4) bradyzoites) of TgCtwh1 isolate of T. gondii isolated from Wuhan, China. Cysts were isolated from the entire brain of mice infected with TgCtwh1 by density gradient centrifugation over Fycoll-paque plus. Animals were orally inoculated with cysts on day zero, and peripheral blood, lymph nodes, heart, liver, and brain of infected mice were collected on days 2, 4, 7, 10, 14, 21, 35, 50, and 72 post infection. Five mice were sacrificed by cervical dislocation under anesthesia at each time of collection, and the kinetic distribution was detected by fluorescence quantitative PCR and tissue inoculation into fresh mice. The cyst formation at various intervals after infection was also observed, as was the number of the cysts in brains and the cyst-forming rate. RESULTS: The body weight of the mice lessened (3.650 +/- 0.252)g post oral infection on day 7, and the weight was progressively decreased between day 10 [(1.730 +/- 0.017)g] and day 14 [(-0.390 +/- 0.554) g] after infection (P<0.05). In the brain tissue, cysts were first observed on day 21 post oral infection and the cyst-forming rate was 80%, and the average diameter of cysts was 20-40 microm. While on day 35 after infection, the cysts were formed in all infected mice(cyst-forming rate was 100%) and the average diameter was 50-60 microm. In chronic infection, DNA copies of parasites were first detected in blood, heart, liver and lymph node at 3.51 +/- 0.152, 4.100 +/- 0.198, 4.220 +/- 0.209 and 4.960 +/- 0.052 respectively on day 2, then in the brain on day 4 (3.800 +/- 0.154). During the early days of infection, the parasite burden in blood was progressively increased until days 7 (5.240 +/- 0.115) then gradually decreased and become undetectable on day 35. The burden of T. gondii in the heart and brain tissues increased significantly and reached their maximum on day 14 (5.640 +/- 0.214) and day 10 (5.790 +/- 0.060), respectively, and remained a stable level thereafter. Liver and lymph tissues reached their maximum on day 7 (5.310 +/- 0.038) and day 10 (6.200 +/- 0.152), then gradually decreased and become undetectable on day 50. CONCLUSION: The parasitemia in mice infected with T. gondii cyst-forming isolate lasts for 21 d at least, and cysts are detected in brain on day 21.