ABSTRACT
Objective To investigate the functions of the KIFC1 gene in tumor cells and its effect on the proliferation of cervical cancer cells. Methods We designed sgRNAs targeting the KIFC1 gene and constructed a recombinant plasmid based on the pSpCas9 (BB)-2A-GFP vector, which was co-transfected into HeLa cells. We screened monoclonal knockout cell lines through flow cytometry sorting, limited dilution inoculation of cells, and sequencing. RT-qPCR, Western blot, and immunofluorescence were used to detect the transcription and protein expression levels of KIFC1 in knockout cells. Cell phenotypes such as nucleus and microtubule cytoskeleton were observed using phase-contrast microscopy and fluorescence confocal microscopy. Cell proliferation, cell cycle, and apoptosis were analyzed by growth curve plotting, EdU labeling, and acridine orange staining. Results The deletion of the KIFC1 gene resulted in the abnormal phenotypes of HeLa cells, with increased numbers of multinuclei, micronucleus, and disordered microtubules. The cell cycle was disrupted, accompanied with a significant increase in the ratio of late apoptotic cells and a decrease in cell proliferation (all P < 0.05). Conclusion KIFC1 gene deletion affects the assembly of microtubules and cell division in HeLa cells, leading to abnormal nuclear morphology, chromatin elimination, cell cycle arrest, and increased cell apoptosis.
ABSTRACT
Objective:To explore the effects of sitagliptin phosphate combined with metformin on blood glucose control and microinflammation in patients with type 2 diabetes.Methods:One hundred patients with newly diagnosed type 2 diabetes who were treated in the First Affiliated Hospital of Xingtai Medical College from March 2017 to March 2019 were randomly divided into observation group (50 cases) and control group (50 cases). The observation group was treated with sitagliptin phosphate combined with metformin for 8 weeks, while the control group was treated with metformin for 8 weeks. The changes of fasting blood-glucose (FBG) and blood glucose 2 h after meal (2 h-PBG ) in the two groups before and after treatment were observed, and the standard time of FBG and 2 h-PBG in the two groups were statistically analyzed. The levels of interleukin(IL)-1, IL-6 and high-sensitivity C-reactive protein (hS-CRP) were compared between the two groups before and after treatment.Results:After treatment, the levels of FBG and 2 h-PBG in the observation group were significantly lower than those in the control group: (6.32 ± 0.83) mmol/L vs. (7.21 ± 1.03) mmol/L, (8.61 ± 1.26) mmol/L vs. (9.63 ± 1.12) mmol/L, and the standard time of FBG and 2 h-PBG in the observation group were significantly shorter than those in the control group: (3.11 ± 0.86) weeks vs. (4.53 ± 1.31) weeks, (3.26 ± 0.36) weeks vs. (9.63 ± 1.12) weeks, and the differences were statisticlly significant ( P<0.05). After treatment, the serum levels of IL-1, IL-6 and hs-CRP in the observation group were significantly lower than those in the control group: (22.86 ± 4.07) ng/L vs. (35.13 ± 5.92) ng/L, (5.93 ± 0.84) ng/L vs. (9.67 ± 1.11) ng/L, (2.12 ± 0.25) ng/L vs. (3.57 ± 0.48) ng/L, and the differences were statistically significants ( P<0.05). Conclusions:Sitagliptin phosphate combined with metformin in the treatment of type 2 diabetes patients can rapidly and effectively control blood glucose and improve the state of microinflammation in patients.