MiR-130a regulating the biological function of colon cancer by targeting inhibition of PTEN.

OBJECTIVE: To investigate the expression of miR-130a in human colon cancer patients and its specific mechanism of regulating the biological function of colon cancer cells. PATIENTS AND METHODS: Cancer tissues, paracancerous tissues, and serum samples of 40 colon cancer patients who underwent surgery in The Second Affiliated Hospital of Qiqihar Medical University from May 2018 to March 2019 were collected, and 40 healthy volunteers who received physical examination in The Second Affiliated Hospital of Qiqihar Medical University were collected. Real Time-quantitative Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of miR-130a. Human colon cancer cell was divided into miR-130a mimic group, miR-130a inhibitor group, mimic NC (negative control), and inhibitor NC group. QRT-PCR was used to detect the expression of miR-130a, and MTT assay, colony formation assay, cell scratch assay, transwell assay were performed to detect cell viability, proliferation, migration, and invasion ability. RESULTS: Compared with adjacent tissues, the expression of miR-130a was significantly increased in colon cancer tissues (p=0.0125); the expression of miR-130a in transfected miR-130a mimic group was higher than that in NC group, but the expression in transfected miR-130a inhibitor group was significantly lower than that in NC group; overexpression of miR-130a significantly increased cell viability, proliferation, migration, and invasion of colon cancer cells, while knockdown of miR-130a significantly inhibited colon cancer cell biological activity; target prediction, qRT-PCR, and Western blot assays showed that miR-130a participated in the development and progression of colon cancer by targeting inhibition of PTEN expression. CONCLUSIONS: The expression of miR-130a in serum and cancer tissues of colon cancer patients is significantly increased, and it can regulate the biological function of colon cancer cells by inhibiting the expression of target gene PTEN. Knockdown of miR-130a may be used as a new clinical treatment for colon cancer.

Icariin inhibits the osteoclast formation induced by RANKL and macrophage-colony stimulating factor in mouse bone marrow culture.

Icariin is a prenylated flavonol glycoside contained in the herb Epimedium, which has long been used to improve bone fracture healing or prevent osteoporosis because of the belief that the herb has bone-strengthening action. We have previously demonstrated that icariin enhances the osteogenic differentiation of rat bone marrow stromal cells, and partially explained the bone-strengthening mechanism of the herb. In the present study, the effect of icariin on osteoclastogenesis and bone resorption activity was investigated in mouse bone marrow culture. It was found that icariin dose-dependently inhibited the growth and differentiation of hemopoietic cells from which osteoclasts were formed. Far less TRAP+ multinuclear cells appeared in the 10 microM icariin group than in the control. The bone resorption pits formed in the 10 microM icariin group was also significantly less than that of the control. RT-PCR analysis showed that the gene expression of TRAP, RANK and CTR was obviously lower than that of the control. It can be concluded that icariin has the ability to inhibit the formation and bone resorption activity of osteoclasts, which suggests that icariin should be the effective component for the bone-strengthening action of herb Epimedium.

Icariin enhances the osteogenic differentiation of bone marrow stromal cells but has no effects on the differentiation of newborn calvarial osteoblasts of rats.

Since the total flavonoid extract (TFE) of Epimedium herb was found to prevent osteoporosis induced by ovariectomy in rats, we have been attempting to identify the exact compound responsible for the bone-strengthening activity. In this experiment, four flavonoid extracts were obtained from Epimedium sagittatum (Siebold & Zucc.) Maxim, which contained 25.3%, 51.2%, 82.3% and 99.2% icariin respectively. They were separately supplemented into the culture media of newborn rat calvarial osteoblasts (ROB) or primary rat bone marrow stroma cells (rMSCs) at 0.1, 1, 10 and 100 microg/ml respectively, in order to observe their effects on the cells. Not any appreciable effect was found on the differentiation of ROB, but an enhancing effect on the osteogenic differentiation of rMSCs was found, and the enhancing degree was icariin-dependent, that is, a higher concentration of icariin in the extract caused more mineralized bone nodules and higher calcium deposition levels. The gene expressions involved in osteogenesis were also improved which was revealed by RT-PCR, including alkaline phosphatase, bone matrix protein (osteocalcin, osteopontin, bone sialoprotein) and cytokines (TGF-beta1 and IGF-I). The effect of icariin on cell proliferation was assayed by the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Icariin inhibited the proliferation of rMSCs and ROB when its concentration was higher than 10(-5) microM (6.7 microg/ml), no stimulative effect was found. The above results indicated that icariin may exert bone-strengthening activity by enhancing the osteogenic differentiation of MSCs, which partially explains the anti-osteoporosis action of Epimedium herb.

Icariin, a flavonoid from the herb Epimedium enhances the osteogenic differentiation of rat primary bone marrow stromal cells.

The herb Epimedium has long been used in Traditional Chinese Medicine to treat bone fracture and prevent osteoporosis. Researchers believe that the flavonoids contained in the herb are the effective component for this activity. However, no single flavonoid has been studied for its effect on bone-related cells. In the present study, icariin, one of the major flavonoids of the herb, supplemented the primary culture medium of rat bone marrow stromal cells (rMSCs) at 0.1 microM , 1 microM and 10 microM respectively. It was found that icariin stimulated the proliferation of rMSCs and increased the number of CFU-F stained positive for alkaline phosphatase in a dose-dependent manner. Icariin also dose-dependently increased the alkaline phosphatase activity, osteoalcin secretion and calcium deposition level of rMSCs during osteogenic induction. The addition of 10 microM icariin caused four times more mineralized bone nodules to be formed by rMSCs than in the control. The results demonstrated that icariin should be an effective component for bone-strengthening activity, and one of the mechanisms is to stimulate the proliferation and enhance the osteogenic differentiation of MSCs.

An implant of a composite of bovine bone morphogenetic protein and plaster of paris for treatment of femoral shaft nonunions.

Sixteen patients with nonunion of femoral shaft fractures were treated by implantation of a composite of bovine bone morphogenetic protein and plaster of Paris (bBMP/PLP). There was an average of 1.5 surgical procedures per patient attempting union prior to bBMP/PLP implantation The implant was prepared in an aggregate of 50 mg bBMP in 1 gram of plaster of Paris. It was implanted directly to fill the fracture gaps and packed around the fracture lines. Seventeen operations were done on the 16 patients. Union was obtained in 16 of the 17 operations. The average time of union for the 16 successful operations was 5.7 months. There were no significant clinically manifested postoperative complications.