To observe the efficacy of low-dose triptorelin acetate injection combined with vitamin D drop in the treatment of central precocious puberty (CPP) in girls.

CPP children were divided into treatment group and control group using a cohort method. The treatment group received a low-dose of triptorelin acetate injection at 50 μg·kg^-1 via intramuscular injection every 4 weeks, along with oral vitamin D drop at 400 units. The control group received standard dose treatment, starting with an initial dose of 100 μg·kg^-1 via intramuscular injection, followed by a maintenance dose of 80 μg·kg^-1 every 4 weeks. Both groups were treated continuously for 12 months. The clinical efficacy, physical development, hormone levels, secretion levels of peptides and hormones related to the hypothalamic-pituitary-gonadal axis were compared between the two groups, along with safety evaluation.

During the trial, 42 cases were included in the treatment group and 37 cases in the control group. After treatment, the overall effective rates for the treatment and control groups were 90.48% (38 cases/42 cases) and 78.38% (29 cases/37 cases), respectively, with no statistically significant difference (P>0.05). After treatment, the predicted adult height for the treatment and control groups was (164.59±4.35) and (162.38±5.21) cm, respectively; the differences in bone age were (1.07±0.28) and (1.24±0.29) years; the growth rates were (5.35±1.17) and (5.93±1.24) cm·year^-1; estradiol levels were (13.11±5.67) and (16.08±4.73) pg·mL^-1; luteinizing hormone levels were (1.49±0.20) and (2.15±0.18) U·L^-1; follicle-stimulating hormone levels were (4.95±0.74) and (6.14±0.87) pg·mL^-1; neuropeptide kisspeptin levels were (1.02±0.19) and (1.20±0.40) μg·L^-1; neurokinin B levels were (0.35±0.08) and (0.41±0.08) μg·L^-1; anti-Müllerian hormone levels were (3.88±0.36) and (4.15±0.40) ng·L^-1; insulin-like growth factor-1 levels were (216.91±25.43) and (231.64±30.87) ng·mL^-1; 25-hydroxyvitamin D levels were (28.42±4.52) and (26.15±3.73) μg·L^-1, with statistically significant difference (all P<0.05). The main adverse drug reactions in the treatment group included nausea and vomiting, as well as pain and swelling at the injection site. In the control group, the main adverse drug reactions included slight vaginal bleeding, nausea and vomiting, pain and swelling at the injection site and rashes near the injection site. The overall incidence of adverse drug reactions in the treatment group and control group were 7.14% (3 cases/42 cases) and 16.22% (6 cases/37 cases), without statistically significant difference (P>0.05).

In children with CPP, the application of low-dose triptorelin combined with vitamin D drop can improve clinical efficacy, physical development, growth-related factors, hormone levels and the secretion levels of peptides and hormones associated with the hypothalamic-pituitary-gonadal axis. It can also slow the development of secondary sexual characteristics and inhibit gonadal development, while demonstrating good safety.

To investigate the inhibition of NG108-15 cells proliferation by Xanthohumol (XN) and its effects on reactive oxygen species (ROS), extracellular signal-regulated kinase (ERK) pathway and cell cycle.

NG108-15 cells were divided into control group (conventional culture), experimental-L group, experimental-M group, experimental-H group (20, 40, and 60 μmol·L^-1 XN), NAC combined with XN experimental group [10 μmol·L^-1 N-acetylcysteine (NAC)+40 μmol·L^-1 XN] and ERK inhabitor combined with XN experimental group (20 μmol·L^-1 ERK inhibitor PD98059+40 μmol·L^-1 XN). Cell proliferation capacity was detected by cell counting kit 8 (CCK-8) and trypan blue staining assay; ROS levels were detected by ROS kit; cell cycle and apoptosis levels were detected by flow cytometry; protein relative expression levels of ERK, p-ERK and Caspase 3 were detected by Western blot.

The cell proliferation activities of the control group and the experimental-L group, experimental-M group and experimental-H group were (99.89±1.52)%, (87.57±6.23)%, (51.96±3.08)% and (37.11±1.57)%, respectively; the cell death rates were (4.65±1.57)%, (15.54±1.94)%, (41.06±2.77)% and (65.22±5.81)%, respectively; the average fluorescence intensities of ROS in the control group, the experimental-M group and the NAC combined with XN experimental group were 176.51±25.17, 716.48±95.15 and 543.80±48.91. The relative expression levels of p-ERK protein in the control group, the experimental-M group, and the ERK inhibitor combined with XN experimental group were 0.54±0.04, 1.13±0.18 and 0.31±0.33; the proportions of G2 phase cells were (15.56±1.43)%, (20.31±2.17)% and (16.14±0.81)%, respectively. Compared with the control group, the above indicators in the experimental-L, -M, -H groups showed statistically significant differences (all P<0.001). Compared with the experimental-M group, the control group and the NAC combined with XN experimental group showed statistically significant differences (P<0.05, P<0.01, P<0.001). Compared with the experimental-M group, the ERK inhibitor combined with XN experimental group showed statistically significant differences (P<0.05, P<0.01, P<0.001).

Xanthohumol demonstrates potent inhibition of NG108-15 cells proliferation, with its molecular mechanism likely involving G2/M phase cell cycle arrest through ROS upregulation and activation of the ERK signaling pathway.

To investigate the regulatory mechanism of plantamajoside (PMS) on the growth and metastasis of non-small cell lung cancer (NSCLC).

A549 cells were divided into control group (normal culture), model group (40 μg·mL^-1 PMS), NC inhibitor group (40 μg·mL^-1 PMS+NC inhibitor), miR-525-5p inhibitor group (40 μg·mL^-1 PMS+miR-525-5p inhibitor), sh-NC group (40 μg·mL^-1 PMS+miR-525-5p inhibitor+sh-NC) and sh-ubiquitin conjugating enzyme E2C (UBE2C) group (40 μg·mL^-1 PMS+miR-525-5p inhibitor+sh-UBE2C). Transwell assay was used to evaluate the invasion ability of A549 cells after different treatments. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay was used to evaluate the apoptosis of A549 cells after different treatments. At the animal level, mice were injected with A549 cell suspension (5×10⁶/100 μL) to construct the NSCLC model. They were randomly divided into model-M group, PMS group (50 mg·kg^-1 PMS), miR-525-5p inhibitor-M group (50 mg·kg^-1 PMS+miR-525-5p inhibitor) and sh-UBE2C-M group (50 mg·kg^-1 PMS+miR-525-5p inhibitor+sh-UBE2C). All groups were treated for 5 days with once a day. The size of the transplanted tumor was measured with a ruler every 7 days, and the transplanted tumor was removed and weighed on the 28th day. The number of Transwell invasions in control group, model group, NC inhibitor group, miR-525-5p inhibitor group, sh-NC group and sh-UBE2C group were 173.62±23.46, 82.05±14.17, 84.69±13.82, 143.48±19.81, 139.57±18.24 and 105.93±16.25, respectively; the apoptosis rates were (22.65±4.01)%, (53.34±8.93)%, (51.46±8.81)%, (37.52±5.86)%, (34.83±5.27)% and (48.75±6.19)%, respectively. Compared in model group and control group, compared in miR-525-5p inhibitor group and NC inhibitor group, compared in sh-UBE2C group and sh-NC group, there were significant differences in the number of Transwell invasion and apoptosis rates (all P<0.05). At the animal level, 28 d tumor volumes of model-M group, PMS group, miR-525-5p inhibitor-M group and sh-UBE2C-M group were (966.30±176.53), (495.70±52.40), (841.10±121.25) and (623.80±105.85) mm³, respectively; the tumor weight on day 28 were (1.43±0.52), (0.75±0.21), (1.32±0.43) and (0.85±0.26) g, respectively. Differences of the tumor volume and weight between PMS group and model-M group, sh-UBE2C-M group and miR-525-5p inhibitor-M group were statistically significant at the 28th day (P<0.01, P<0.001).

PMS can significantly inhibit the growth and metastasis of NSCLC by regulating the miR-525-5p/UBE2C axis.

To explore the effects of niraparib (Nira) on ovarian cancer cell proliferation and migration by inhibiting the wingless MMTV integration site family member (Wnt)/beta-catenin (β-catenin) signaling pathway through the regulation of transcription factor 4 gene (TCF4).

SKOV3 cells were divided into control group (normal culture without any treatment), OE-NC group (10 μmol·L^-1 Nira+ transfection OE-NC treatment) and OE-TCF4 group (10 μmol·L^-1 Nira+ transfection OE-TCF4 treatment), experimental-L, M, H groups (5, 10, 15 μmol·L^-1 Nira treatment). Real-time fluorescence quantitative polymerase chain reaction and Western blotting were used to detect the protein and mRNA expression level of TCF4; cell viability was detected by cell counting kit-8 (CCK-8); and cell invasion ability was detected by Transwell. The expression levels of Wnt/β-catenin pathway-related proteins were detected by immunofluorescence.

The 48 h cell survival rates in control group, experimental-L, -M, -H groups were (98.96±3.13)%, (77.53±7.03)%, (58.31±7.52)% and (35.88±5.97)%, respectively. The relative mRNA expression levels of TCF4 in the control group and the experimental-M group were 1.00±0.16 and 0.45±0.07, respectively; while the relative protein expression levels of TCF4 were 1.00±0.12 and 0.43±0.06, respectively. The number of invasive cells in control group, experimental-M group, OE-NC group and OE-TCF4 group were 115.77±16.57, 60.81±8.80, 66.43±8.98 and 99.25±13.73, respectively; the relative fluorescence intensities of β-catenin were 1.00±0.13, 0.15±0.03, 0.17±0.04 and 0.60±0.11; the relative fluorescence intensities of cancer-Myc were 1.00±0.15, 0.07±0.01, 0.08±0.02 and 0.54±0.09, respectively; the relative fluorescence intensities of matrix metalloproteinase-9 (MMP-9) were 1.00±0.13, 0.06±0.01, 0.07±0.02 and 0.28±0.05, respectively. The control group was compared with experimental-M group, and the OE-TCF4 group was compared with OE-NC group, there were statistically significant differences in the above indexes (all P<0.05).

Nira can inhibit the activity and invasion of ovarian cancer SKOV3 cells, which may be related to the down-regulation of TCF4 to inhibit the activation of Wnt/β-catenin signaling pathway.

To explore the specific mechanism of the gastric protective effect of salvianolic acid B (Sal B) on precancerous lesions of the stomach (PLGC) in rats by regulating the microRNA (miR)-106b-5p/B-cell translocation gene 3 (BTG3) molecular axis.

Except for the NC group, the rat models of PLGC were established. The rats were randomly divided into the NC group (without any treatment, intragastric administration of normal saline), the model group (after successful modeling, intragastric administration of normal saline), the experimental group (intragastric administration of Sal B based on the model group), the NC mimic group (based on the experimental group, injection of NC mimic plasmid via the tail vein), the miR-106b-5p mimic group (based on the experimental group, injection of miR-106b-5p mimic plasmid via the tail vein), the pcDNA3.1-NC group (based on the miR-106b-5p mimic group, intraperitoneal injection of pcDNA3.1-NC plasmid) and the pcDNA3.1-BTG3 group (based on the miR-106b-5p mimic group, intraperitoneal injection of pcDNA3.1-BTG3 plasmid). Quantitative real-time polymerase chain reaction experiments were carried out to detect the mRNA expression levels of miR-106b-5p, BTG3 and inflammatory factors in the gastric mucosa tissues. Enzyme-linked immunosorbent assays were performed to detect the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA) and gastrin-17 (G-17) in the serum. Western blot experiments were used to detect the levels of proteins related to the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) signaling pathway and mucin 2 (MUC2) in the gastric mucosa tissues.

The relative expression levels of miR-106b-5p in the NC group, the model group and the experimental group were 1.00±0.08, 1.76±0.35 and 1.33±0.26, respectively, and the relative expression levels of BTG3 mRNA were 1.00±0.05, 0.39±0.06 and 0.81±0.15, respectively. There were statistically significant differences in the above-mentioned indicators between the model group and the NC group, as well as between the experimental group and the model group (all P<0.05). The levels of SOD in the NC group, the model group, the experimental group, the NC mimic group, the miR-106b-5p mimic group, the pcDNA3.1-NC group and the pcDNA3.1-BTG3 group were (197.81±47.15), (118.54±20.49), (172.29±34.34), (176.78±35.57), (127.65±21.23), (119.41±20.50) and (158.28±31.07) U·mL^-1, respectively; the levels of MDA were (3.44±0.67), (8.08±1.32), (3.92±0.75), (3.84±0.70), (6.50±1.21), (6.34±1.17) and (4.22±0.79) nmol·mL^-1, respectively; the levels of G-17 were (221.57±40.26), (155.62±28.43), (201.13±37.35), (194.85±35.24), (162.72±31.65), (160.04±29.37) and (189.32±32.15) pg·mL^-1, respectively; the levels of PI3K were 1.00±0.14, 1.81±0.36, 1.23±0.21, 1.19±0.20, 1.65±0.32, 1.70±0.35 and 1.38±0.28, respectively; the ratios of phosphorylated AKT/AKT were 1.00±0.09, 1.94±0.39, 1.27±0.23, 1.21±0.18, 1.77±0.37, 1.73±0.33 and 1.42±0.26, respectively. There were statistically significant differences in the above-mentioned indicators between the miR-106b-5p mimic group and the NC mimic group, and between the pcDNA3.1-BTG3 group and the pcDNA3.1-NC group (all P<0.05).

Sal B may exert a gastric protective effect by down-regulating the targeted negative regulation of BTG3 expression by miR-106b-5p, inhibiting the activation of the PI3K/AKT signaling pathway, thereby alleviating the oxidative stress and inflammatory response in PLGC rats and improving the condition of the gastric mucosa.

To observe the clinical efficacy and safety of agomelatine combined with paroxetine tablets in patients with depression and insomnia, and its effects on neurotransmitter levels and sleep.

The patients with depression and insomnia were divided into control group and treatment group, according to random number table. Control group was treated with paroxetine tablets 20 mg, oral, treatment course was 8 weeks. Treatment group was treated with agomelatine 25 mg and paroxetine tablets 20 mg, once a day, treatment course was 8 weeks. The Hamilton depression rating scale (HAMD), Montreal cognitive assessment scale (MoCA), neurotransmitters [norepinephrine (NE), 5-hydroxytryptamine (5-HT), dopamine (DA)], Pittsburgh sleep quality index (PSQI) and incidence of adverse drug reactions between two groups were compared.

Treatment group was enrolled 60 cases; control group was enrolled 60 cases. After 8 weeks of treatment, the HAMD in treatment group and control group were (17.33±1.54) and (21.19±1.87) scores; MoCA were (29.01±0.19) and (28.52±0.21) scores; NE levels were (65.56±4.63) and (53.74±4.52) ng·mL^-1; 5-HT levels were (165.82±7.17) and (149.63±8.48) ng·mL^-1; DA levels were (582.73±17.52) and (543.85±20.38) ng·mL^-1, sleep latency were (1.09±0.12) and (1.53±0.15) scores; sleep efficiency were (1.12±0.17) and (1.58±0.20) scores; hypnotic drug were (1.01±0.13) and (1.42±0.15) scores; subjective sleep quality were (1.14±0.10) and (1.62±0.16) scorse; sleep persistence were (1.16±0.11) and (1.55±0.13) scores; the score for sleep disorders were (1.07±0.14) and (1.52±0.17) scores; daytime functional disorders were (0.99±0.16) and (1.51±0.18) scores, all with statistically significant difference (all P<0.05). The adverse drug reactions in treatment group and control group were 11.67% and 8.33%, with no statistically significant differences (P>0.05).

Administration of agomelatine and paroxetine tablets to patients with depression and insomnia can improve depressive symptoms, cognitive function, sleep conditions, regulate the expression of neurotransmitters such as NE and 5-HT, and has good safety.

To investigate the effects of Cepharanthine on apoptosis, energy metabolism and angiogenesis of breast cancer cells by regulating the microRNA (miR)-378c/Golgi transport 1A protein (GOLT1A) axis.

MDA-MB-231 cells in the logarithmic growth phase were divided into the following groups: Control group (no treatment), NC inhibitor group (transfected with miR-378c inhibitor negative control), miR-378c inhibitor group (transfected with miR-378c inhibitor), si-NC group (transfected with si-NC plasmid), si-GOLT1A group (transfected with si-GOLT1A plasmid), Cepharanthine group (20 μmol·L^-1 Cepharanthine), Cepharanthine combined with miR-378c inhibitor group (Cepharanthine group with miR-378c inhibitor transfection) and miR-378c inhibitor combined with si-GOLT1A group (Cepharanthine group with co-transfection of miR-378c inhibitor and si-GOLT1A plasmid). Real-time fluorescence quantitative polymerase chain reaction was used to detect the relative expression levels of miR-378c, GOLT1A, vascular endothelial growth factor A (VEGFA) and thrombospondin-1 (THBS1); kits were used to detect energy metabolism indicators; immunofluorescence was used to detect VEGFA and THBS1 expression levels.

The relative expression levels of miR-378c in the control group and Cepharanthine group were 1.00±0.14 and 3.26±0.48, respectively; the relative expression levels of GOLT1A mRNA were 1.00±0.15 and 0.43±0.07, respectively. The glutamine concentrations in the control group, Cepharanthine group, Cepharanthine combined with miR-378c inhibitor group and miR-378c inhibitor combined with si-GOLT1A group were (22.03±3.14), (8.27±1.94), (17.25±2.86) and (13.11±2.28) nmol·10^-7 cells, respectively; the acetyl coenzyme A concentrations were (11.32±2.06), (5.71±1.19), (9.62±1.85) and (6.67±1.63) nmol·10^-6 cells, respectively; the succinate concentrations were (53.67±6.92), (21.24±3.87), (39.82±4.39) and (30.36±4.21) nmol·mg^-1·Pr^-1, respectively; the ATP production values were (2.32±0.42), (0.96±0.12), (1.91±0.33) and (1.16±0.26) mmol·mg^-1·Pr^-1, respectively; the relative expression levels of VEGFA mRNA were 1.00±0.14, 0.56±0.10, 0.84±0.13 and 0.69±0.10, respectively; the relative expression levels of THBS1 mRNA were 1.00±0.12, 3.43±0.57, 2.04±0.36 and 3.10±0.52, respectively; the relative fluorescence intensities of VEGFA were 1.00±0.11, 0.37±0.05, 0.76±0.09 and 0.48±0.06, respectively; and the relative fluorescence intensities of THBS1 were 1.00±0.09, 3.71±0.62, 1.88±0.31 and 2.55±0.37, respectively. All the above indicators showed significant differences between the control group and the Cepharanthine group, the Cepharanthine group and the Cepharanthine combined with miR-378c inhibitor group, the Cepharanthine combined with miR-378c inhibitor group and the miR-378c inhibitor combined with si-GOLT1A group (P<0.05, P<0.01, P<0.001).

Cepharanthine can promote apoptosis and inhibit energy metabolism and angiogenesis in breast cancer cells, which may be related to its regulation of the miR-378c/GOLT1A axis.

To investigate the effect of Rocaglamide (Roc-A) on paclitaxel (PTX) resistance in breast cancer MCF-7/PTX cell line and its mechanism.

With breast cancer MCF-7 cells as the research object, PTX resistant MCF-7/PTX cell line was obtained by continuous induction with low concentration of PTX. Methyl thiazolyl tetrazolium (MTT) method was used to determine the drug resistance index of MCF-7/PTX cell line, the effect of Roc-A on the proliferation activity of MCF-7/PTX cell line and the drug resistance reversal multiple. MCF-7/PTX cell lines were divided into control group (normal cultured cells), PTX group (40.8 nmol·L^-1 PTX), PTX+Roc-A-L group (40.8 nmol·L^-1 PTX+25 nmol·L^-1 Roc-A), PTX+Roc-A-M group (40.8 nmol·L^-1 PTX+50 nmol·L^-1 Roc-A) and PTX+Roc-A-H group (40.8 nmol·L^-1 PTX+100 nmol·L^-1 Roc-A). The apoptosis level of each group was detected by flow cytometry; the relative expression levels of cell autophagy related protein LC3 were detected by immunofluorescence assay; Western blotting was used to detect the relative expression levels of cell apoptosis related proteins, such as B cell lymphoma-2 associated X protein (Bax) and autophagy associated protein autophagy microtubule associated protein light chains 3 (LC3).

The PTX resistant cell line MCF-7/PTX of breast cancer was successfully constructed with a drug resistance index of 5.49; Roc-A could inhibit the proliferation activity of MCF-7/PTX cell lines in a concentration dependent manner, while the reversal multiples of PTX resistance of MCF-7/PTX cell lines to Roc-A at different concentrations (25, 50, 100 nmol·L^-1) were 1.57, 2.68 and 6.16, respectively. The apoptosis rates of the control group, PTX group, PTX+Roc-A-L group, PTX+Roc-A-M group and PTX+Roc-A-H group were (4.75±2.10)%, (21.79±3.61)%, (32.54±3.43)%, (39.82±3.26)% and (55.61±4.22)%, respectively; the relative expression levels of LC3 fluorescence were 1.00±0.08, 0.77±0.06, 0.60±0.06, 0.41±0.04 and 0.26±0.03, respectively; the relative expression levels of Bax protein were 0.12±0.04, 0.25±0.09, 0.48±0.10, 0.86±0.12 and 1.10±0.13, respectively; the LC3Ⅱ/LC3Ⅰ protein ratio were 6.52±0.27, 3.75±0.23, 1.76±0.17, 1.31±0.10 and 1.02±0.06, respectively. Compared with the control group, the PTX group, PTX+Roc-A-L group, PTX+Roc-A-M group, PTX+Roc-A-H group showed statistically significant differences in the above indicators (all P<0.05); the above indicators in the PTX+Roc-A-L group, PTX+Roc-A-M group and PTX+Roc-A-H group were compared with those in the PTX group, and the differences were statistically significant (all P<0.05), showing a dose-dependent relationship.

Roc-A can reverse the resistance of breast cancer drug resistant cell line MCF-7/PTX to PTX, and the mechanism may be related to the reduction of autophagy.

To establish a qualification evaluation system for primary investigators (PI) in clinical trials.

Based on the preliminary determination of PI qualification evaluation indicators in accordance with laws and regulations, 19 experts engaged in clinical trials or clinical trial management fields were selected for 2 rounds of Delphi expert consultation, and the weights of each indicator were calculated using principal component analysis.

The positive coefficients for the two rounds of expert consultation were 100.00% and 94.74%, and the expert authority coefficients were 0.940 and 0.933, the coordination coefficients of expert opinions were 0.318 and 0.613. After 2 rounds of expert consultation, the PI qualification evaluation indicators were determined, including 3 primary indicators and 17 secondary indicators. The results of principal component analysis showed that the weight range of each index was 0.037 9 to 0.080 3.

The preliminary construction of the PI qualification evaluation system based on the "PI Qualification Evaluation Form" has strong scientificity and applicability, and can provide references for the evaluation and selection of clinical trial PI.

To achieve overexpression of phosphoglycerate kinase 1 (PGK1) in HEK-293T cells and screen the key site of its succinylation through constructing PGK1 overexpression vector (PGK1-WT) and four lysine site succinylation mutation vectors.

GEPIA2 database was used to analyze the expression of PGK1 mRNA in normal breast tissue (291 cases) and breast cancer tissue (135 cases) and the relationship between PGK1 expression level and the overall survival rate of breast cancer patients. Protein modification proteomics was used to detect the levels of protein succinylation in four triple negative breast cancer (TNBC) cell lines (MDA-MB-436, MDA-MB-468, MDA-MB-231, BT-549) and normal breast epithelial cells (MCF-10A); the succinylated positive and negative mutation vectors of K11, K131, K323, K361 were contructed with point mutation kit. The experiment was divided into NC, shPGK1-496, shPGK1-220, PGK1-WT groups and positive and negative mutant groups of K11, K131, K323 and K361. Real-time fluorescence quantitative polymerase chain reaction was used to detect the level of expression of PGK1 mRNA in NC, shPGK1-496 and shPGK1-220 groups. The expression levels of PGK1 proteins in NC, shPGK1-496 and shPGK1-220 groups, and the expression levels of Flag proteins in NC, PGK1-WT and four sites in positive and negative mutant groups were detected by Western blot. The key site of PGK1 succinylation was detected by co-immunoprecipitation.

Compared with normal breast tissues, the level of PGK1 mRNA was highly expressed in breast cancer tissue, and the survival time of breast cancer patients with high PGK1 expression was shorter. Four lysine sites of PGK1 in TNBC cells were hypersuccinylated. The relative expression levels of PGK1 mRNA in NC, shPGK1-496 and shPGK1-220 groups were 1.03±0.23, 0.15±0.11 and 1.42±0.69, respectively; the relative expression levels of PGK1 protein were 1.08±0.05, 0.49±0.05 and 1.28±0.01 respectively; compared with NC group, the above indexes in shPGK1-496 group were statistically different (all P<0.01). The relative expression levels of Flag protein in NC group, PGK1-WT group and positive and negative mutant groups of K11, K131, K323 site were 0, 3.43±0.09, 3.28±0.10, 2.28±0.11, 3.03±0.03, 2.79±0.11, 3.29±0.09 and 2.98±0.04, respectively; compared with the NC group, PGK1-WT group and the positive and negative mutant groups of K11, K131 and K323 had statistical differences (all P<0.01). The relative expression levels of Flag protein in NC group, PGK1-WT group and K361 positive and negative mutant group were 0, 2.58±0.11, 3.04±0.02 and 4.04±0.10, respectively; compared with NC group, PGK1-WT group and K361 positive and negative mutant group had statistical significance (all P<0.01). The levels of lysine succinylation in IgG group, PGK1-WT group and negative mutant groups of K11, K131, K323, K361 were 0, 1.53±0.12, 1.30±0.18, 1.29±0.09, 1.23±0.03 and 0.89±0.16, respectively; compared with PGK1-WT group, the negativ mutant group of K361 had statistical significance (P<0.01).

The four lysine sites of PGK1 in TNBC cells are highly succinylated, and K361 site is critical to their succinylation.